#### Sample records for cd-sensitive zn-porphyrin tweezer

1. Analysis of the nitrogen K-edge x-ray absorption spectra of Zn-porphyrin/C{sub 70}-fulleren complex for solar cells

Suchkova, S A; Soldatov, A [Southern Federal University, Sorge str. 5, 344090 Rostov-on-Don (Russian Federation); Cudia, C Castellarin, E-mail: suchkova_sv@inbox.r [Sincrotrone Trieste S.C.p.A, s.s. 14 km 163.5 in Area Science Park, 34012 Trieste (Italy)

2009-11-15

The atomic structure models of Zn-porphyrin/C{sub 70} multilayer for solar cells were examined. The local atomic structure of the Zn-porphyrin/C{sub 70} complex was refined with the use of previously published results [1]. Since near-edge spectral region (XANES) is sensitive to the three-dimensional atomic geometry, the theoretical analysis of the experimental XANES was performed on the basis of finite difference method (FDMnes 2008 program code). Some electronic properties of the complex were obtained from the DFT calculations performed by means of Amsterdam Density Functional program package.

2. Compact laser tweezers

Šerý, Mojmír; Lošťák, Z.; Kalman, M.; Jákl, Petr; Zemánek, Pavel

Bellingham: SPIE, 2007, 6609:1-7. ISBN 978-0-8194-6748-5. ISSN 0277-786X. [Czech-Polish-Slovak Conference Wave and Quantum Aspects of Contemporary Optics /15./. Liberec (CZ), 11.09.2006-15.09.2006] R&D Projects: GA MPO FT-TA2/059 Institutional research plan: CEZ:AV0Z20650511 Keywords : laser tweezers * laser diode * optical manipulation Subject RIV: BH - Optics, Masers, Lasers

3. Optical tweezers absolute calibration

Dutra, R S; Neto, P A Maia; Nussenzveig, H M

2014-01-01

Optical tweezers are highly versatile laser traps for neutral microparticles, with fundamental applications in physics and in single molecule cell biology. Force measurements are performed by converting the stiffness response to displacement of trapped transparent microspheres, employed as force transducers. Usually, calibration is indirect, by comparison with fluid drag forces. This can lead to discrepancies by sizable factors. Progress achieved in a program aiming at absolute calibration, conducted over the past fifteen years, is briefly reviewed. Here we overcome its last major obstacle, a theoretical overestimation of the peak stiffness, within the most employed range for applications, and we perform experimental validation. The discrepancy is traced to the effect of primary aberrations of the optical system, which are now included in the theory. All required experimental parameters are readily accessible. Astigmatism, the dominant effect, is measured by analyzing reflected images of the focused laser spo...

4. Optical tweezers principles and applications

Jones, Philip; Volpe, Giovanni

2015-01-01

Combining state-of-the-art research with a strong pedagogic approach, this text provides a detailed and complete guide to the theory, practice and applications of optical tweezers. In-depth derivation of the theory of optical trapping and numerical modelling of optical forces are supported by a complete step-by-step design and construction guide for building optical tweezers, with detailed tutorials on collecting and analysing data. Also included are comprehensive reviews of optical tweezers research in fields ranging from cell biology to quantum physics. Featuring numerous exercises and problems throughout, this is an ideal self-contained learning package for advanced lecture and laboratory courses, and an invaluable guide to practitioners wanting to enter the field of optical manipulation. The text is supplemented by www.opticaltweezers.org, a forum for discussion and a source of additional material including free-to-download, customisable research-grade software (OTS) for calculation of optical forces, dig...

5. Coaxial Atomic Force Microscope Tweezers

Brown, K A; Westervelt, R M

2010-01-01

We demonstrate coaxial atomic force microscope (AFM) tweezers that can trap and place small objects using dielectrophoresis (DEP). An attractive force is generated at the tip of a coaxial AFM probe by applying a radio frequency voltage between the center conductor and a grounded shield; the origin of the force is found to be DEP by measuring the pull-off force vs. applied voltage. We show that the coaxial AFM tweezers (CAT) can perform three dimensional assembly by picking up a specified silica microsphere, imaging with the microsphere at the end of the tip, and placing it at a target destination.

6. Coaxial atomic force microscope tweezers

Brown, K. A.; Aguilar, J. A.; Westervelt, R. M.

2010-03-01

We demonstrate coaxial atomic force microscope (AFM) tweezers that can trap and place small objects using dielectrophoresis (DEP). An attractive force is generated at the tip of a coaxial AFM probe by applying a radio frequency voltage between the center conductor and a grounded shield; the origin of the force is found to be DEP by measuring the pull-off force versus applied voltage. We show that the coaxial AFM tweezers can perform three-dimensional assembly by picking up a specified silica microsphere, imaging with the microsphere at the end of the tip, and placing it at a target destination.

7. Multi-Beam Optical Tweezers

2003-01-01

A set of multi-beam electromagnetic tweezers is provided comprising a multi-beam generator for emission of a plurality of electromagnetic beams, at least some of the electromagnetic beams intersecting each other, or, having an individually controlled polarization whereby the position and/or angul...

8. Optical tweezers technique and its applications

Guo, HongLian; Li, ZhiYuan

2013-12-01

Since their advent in the 1980s, optical tweezers have attracted more and more attention due to their unique non-contact and non-invasion characteristics and their wide applications in physics, biology, chemistry, medical science and nanoscience. In this paper, we introduce the basic principle, the history and typical applications of optical tweezers and review our recent experimental works on the development and application of optical tweezers technique. We will discuss in detail several technological issues, including high precision displacement and force measurement in single-trap and dual-trap optical tweezers, multi-trap optical tweezers with each trap independently and freely controlled by means of space light modulator, and incorporation of cylindrical vector optical beams to build diversified optical tweezers beyond the conventional Gaussian-beam optical tweezers. We will address the application of these optical tweezers techniques to study biophysical problems such as mechanical deformation of cell membrane and binding energy between plant microtubule and microtubule associated proteins. Finally we present application of the optical tweezers technique for trapping, transporting, and patterning of metallic nanoparticles, which can be harnessed to manipulate surface plasmon resonance properties of these nanoparticles.

9. Photonic Nanojet in Optical Tweezers

Neves, Antonio Alvaro Ranha

2015-01-01

Photonic nanojets has been brought into attention ten years ago for potential application as ultramicroscopy technique, using its sub-wavelength resolution to enhance detection and interaction with matter. For these novel applications under development, optically trapping a sphere, acts as an ideal framework to employ these nanojets. In this case, the nanojet is generated by a highly focused incident beam contrary to the traditional plane wave. It inherits the advantage from optical trapping, with the microsphere in equilibrium on the beam propagation axis, and be positioned arbitrarily in space, especially for intracellular applications. Moreover, due to optical scattering forces, when in equilibrium, there is a shift of the sphere centre with respect to the beam focus. However, within the stable equilibrium of an optical tweezers configuration, it does not allow the formation of a photonic nanojet. To overcome this, a double optical tweezers, in an unorthodox configuration of two collinearly and co-propagat...

10. Optoelectronic tweezers for medical diagnostics

Kremer, Clemens; Neale, Steven; Menachery, Anoop; Barrett, Mike; Cooper, Jonathan M.

2012-01-01

Optoelectronic tweezers (OET) allows the spatial patterning of electric fields through selected illumination of a photoconductive surface. This enables the manipulation of micro particles and cells by creating non-uniform electrical fields that then produce dielectrophoretic (DEP) forces. The DEP responses of cells differ and can produce negative or positive (repelled or attracted to areas of high electric field) forces. Therefore OET can be used to manipulate individual cells and separate different cell types from each other. Thus OET has many applications for medical diagnostics, demonstrated here with work towards diagnosing Human African Trypanosomiasis, also known as sleeping sickness.

11. Dynamical stabilisation in optical tweezers

Jones, Philip H.; Richards, Christopher J.; Smart, Thomas J.; Cubero, David

2015-03-01

We present a study of dynamical stabilisation of an overdamped, microscopic pendulum realised using optical tweezers. We first derive an analytical expression for the equilibrium dynamically stabilised pendulum position in a regime of high damping and high modulation frequency of the pendulum pivot. This model implies a threshold behavior for stabilisation to occur, and a continuous evolution of the angular position which, unlike the underdamped case, does not reach the fully inverted position. We then test the theoretical predictions using an optically trapped microparticle subject to fluid drag force, finding reasonable agreement with the threshold and equilibrium behavior at high modulation amplitude. Analytical theory and experiments are complemented by Brownian motion simulations.

12. Photonic nanojets in optical tweezers

Photonic nanojets have been brought into attention ten years ago for potential application in ultramicroscopy, because of its sub-wavelength resolution that can enhance detection and interaction with matter. For these novel applications under development, the optical trapping of a sphere acts as an ideal framework to employ photonic nanojets. In the present study, we generated nanojets by using a highly focused incident beam, in contrast to traditional plane waves. The method inherits the advantage of optical trapping, especially for intracellular applications, with the microsphere in equilibrium on the beam propagation axis and positioned arbitrarily in space. Moreover, owing to optical scattering forces, when the sphere is in equilibrium, its center shifts with respect to the focal point of the incident beam. However, when the system is in stable equilibrium with a configuration involving optical tweezers, photonic nanojets cannot be formed. To overcome this issue, we employed double optical tweezers in an unorthodox configuration involving two collinear and co-propagating beams, the precise positioning of which would turn on/off the photonic nanojets, thereby improving the applicability of photonic nanojets. - Highlights: • Photonic nanojets from an optically trapped microsphere are presented. • Electromagnetic beams are described by using beam shape coefficients. • Beam shape coefficients are determined by generalized Lorenz–Mie theory. • Scheme to turn on and off a photonic nanojet is described

13. Ultrafast charge transfer and radiationless relaxations from higher excited state (S2) of directly linked Zn-porphyrin (ZP)-acceptor dyads: investigations into fundamental problems of exciplex chemistry

We have investigated photoinduced electron transfer and related processes from the higher excited electronic state (S2) of Zn-porphyrin-imide acceptor directly linked supramolecular systems (ZP-I) designed especially for the critical studies of the energy gap law (EGL) of the charge separation (CS) from the S2 state and solvent effects upon EGL. We have confirmed the modification of the EGL by change of solvent polarity from acetonitrile (ACN), tetrahydrofuran (THF) to toluene (Tol) and methyl-cyclohexane (MCH), from rather typical bell-shaped one in ACN to that with less prominent normal region and prominent inverted region with moderate slope extending over wider range of -ΔGCS values in nonpolar solvent MCH. We have demonstrated that these solvent effects upon EGL affect delicately various radiationless relaxation processes from S2 state. We have examined also effects of the hydrogen bonding solvent ethanol (EtOH) on the EGL for CS and found very specific effect controlling the CS reaction and related processes

14. Characterising Conical Refraction Optical Tweezers

McDonald, Craig; Rafailov, Edik; McGloin, David

2014-01-01

Conical refraction occurs when a beam of light travels through an appropriately cut biaxial crystal. By focussing the conically refracted beam through a high numerical aperture microscope objective, conical refraction optical tweezers can be created, allowing for particle manipulation in both Raman spots and in the Lloyd/Poggendorff rings. We present a thorough quantification of the trapping properties of such a beam, focussing on the trap stiffness and how this varies with trap power and trapped particle location. We show that the lower Raman spot can be thought of as a single-beam optical gradient force trap, while radiation pressure dominates in the upper Raman spot, leading to optical levitation rather than trapping. Particles in the Lloyd/Poggendorff rings experience a lower trap stiffness than particles in the lower Raman spot but benefit from rotational control.

15. Biaxial crystal-based optical tweezers

Angelsky, Oleg V.; Maksimyak, Andrew P.; Maksimyak, Peter P.;

2010-01-01

We suggest an optical tweezer setup based on an optically biaxial crystal. To control movements of opaque particles, we use shifts. The results of experimental studies are reported which are concerned with this laser tweezer setup. We demonstrate a movement of microparticles of toner using a sing...... singular-optical trap, rotation of particles due to orbital angular momentum of the field, and converging or diverging of two different traps when changing transmission plane of polariser at the input of our polarisation interferometer....

16. Simulation of superresolution holography for optical tweezers

Thomson, L. C.; Boissel, Y.; Whyte, G; Yao, E; Courtial, J.

2008-01-01

Optical tweezers manipulate microscopic particles using foci of light beams. Their performance is therefore limited by diffraction. Using computer simulations of a model system, we investigate the application of superresolution holography for two-dimensional (2D) light shaping in optical tweezers, which can beat the diffraction limit. We use the direct-search and Gerchberg algorithms to shape the center of a light beam into one or two bright spots; we do not constrain the remainder of the bea...

17. Quantitative modeling of forces in electromagnetic tweezers

Bijamov, Alex; Shubitidze, Fridon; Oliver, Piercen M.; Vezenov, Dmitri V.

2010-01-01

This paper discusses numerical simulations of the magnetic field produced by an electromagnet for generation of forces on superparamagnetic microspheres used in manipulation of single molecules or cells. Single molecule force spectroscopy based on magnetic tweezers can be used in applications that require parallel readout of biopolymer stretching or biomolecular binding. The magnetic tweezers exert forces on the surface-immobilized macromolecule by pulling a magnetic bead attached to the free...

18. Holographic Raman Tweezers Controlled by Hand Gestures and Voice Commands

Tomori, Z.; Antalík, M.; Kesa, P.; Kaňka, Jan; Jákl, Petr; Šerý, Mojmír; Bernatová, Silvie; Zemánek, Pavel

2013-01-01

Roč. 3, 2B (2013), s. 331-336. ISSN 2160-8881 Institutional support: RVO:68081731 Keywords : Holographic Optical Tweezers * Raman Tweezers * Natural User Interface * Leap Motion * Gesture Camera Subject RIV: BH - Optics, Masers, Lasers

19. Optical tweezers: light for manipulating microscopic world

Optical tweezers make use of a tightly focused laser beam to trap, move, guide, rotate and even sort microscopic objects solely with light. Although the basic laser tweezers, making use of a TEM00 laser beam to create a single trap point, have proved to be useful for any applications in areas ranging from physics to biology, a major breakthrough in this field came as the use of computer generated holograms enabled researchers to create multiple trap sites from single laser source (holographic optical tweezers). Coupled with microfluidic techniques, holographic optical tweezers have promised development of optical techniques for high throughput sorting of different cell types under a single micro-chip platform. The holographic methods have also helped the use of specialized laser beams like Laguerre-Gaussian beams instead of the conventional laser beam for interesting applications like orienting/rotating the trapped objects or trapping cells with minimum photodamage. Further, combining optical tweezers with Raman spectroscopy is becoming increasingly popular for studying single cell biochemistry as use of optical forces to immobilize the cells under investigations not only avoids the negative effects of fixing the cells onto substrate but also improve the quality of the recorded spectra. These advanced optical trapping techniques as outlined above along with some illustrative biophotonics applications have been explored.

20. Simulation of superresolution holography for optical tweezers

Optical tweezers manipulate microscopic particles using foci of light beams. Their performance is therefore limited by diffraction. Using computer simulations of a model system, we investigate the application of superresolution holography for two-dimensional (2D) light shaping in optical tweezers, which can beat the diffraction limit. We use the direct-search and Gerchberg algorithms to shape the center of a light beam into one or two bright spots; we do not constrain the remainder of the beam. We demonstrate that superresolution algorithms can significantly improve the normalized stiffness of an optical trap and the minimum separation at which neighboring traps can be resolved. We also test if such algorithms can be used interactively, as is desirable in optical tweezers

1. Tomographic phase microscopy using optical tweezers

Habaza, Mor; Gilboa, Barak; Roichman, Yael; Shaked, Natan T.

2015-07-01

We review our technique for tomographic phase microscopy with optical tweezers [1]. This tomographic phase microscopy approach enables full 3-D refractive-index reconstruction. Tomographic phase microscopy measures quantitatively the 3- D distribution of refractive-index in biological cells. We integrated our external interferometric module with holographic optical tweezers for obtaining quantitative phase maps of biological samples from a wide range of angles. The close-tocommon- path, off-axis interferometric system enables a full-rotation tomographic acquisition of a single cell using holographic optical tweezers for trapping and manipulating with a desired array of traps, while acquiring phase information of a single cell from all different angles and maintaining the native surrounding medium. We experimentally demonstrated two reconstruction algorithms: the filtered back-projection method and the Fourier diffraction method for 3-D refractive index imaging of yeast cells.

2. Electric tweezers: negative dielectrophoretic multiple particle positioning

Electric tweezers are a touchless positioning apparatus that employs dielectrophoresis and electroorientation to arbitrarily position cell-sized particles. In this paper, we develop an algorithm which enables electric tweezers to operate on multiple particles. Furthermore, we probe the limits of this technique in simulation, examining the range of electric field magnitudes and forces that can be applied. We then demonstrate this new functionality on two particles. The device can apply forces on any particle of non-zero polarizability and here this is highlighted by manipulating negatively polarized glass beads. Additionally, we demonstrate that negligibly polarized particles can also be manipulated through mechanical forces applied by other particles. (paper)

3. Origin and Future of Plasmonic Optical Tweezers

Jer-Shing Huang

2015-06-01

Full Text Available Plasmonic optical tweezers can overcome the diffraction limits of conventional optical tweezers and enable the trapping of nanoscale objects. Extension of the trapping and manipulation of nanoscale objects with nanometer position precision opens up unprecedented opportunities for applications in the fields of biology, chemistry and statistical and atomic physics. Potential applications include direct molecular manipulation, lab-on-a-chip applications for viruses and vesicles and the study of nanoscale transport. This paper reviews the recent research progress and development bottlenecks and provides an overview of possible future directions in this field.

4. Single Chromatin Fibre Assembly Using Optical Tweezers

Bennink, M.L.; Pope, L.H.; Leuba, S.H.; Grooth, de B.G.; Greve, J.

2001-01-01

Here we observe the formation of a single chromatin fibre using optical tweezers. A single -DNA molecule was suspended between two micron-sized beads, one held by a micropipette and the other in an optical trap. The constrained DNA molecule was incubated with Xenopus laevis egg extract in order to r

5. Quantum computation architecture using optical tweezers

Weitenberg, Christof; Kuhr, Stefan; Mølmer, Klaus;

2011-01-01

We present a complete architecture for scalable quantum computation with ultracold atoms in optical lattices using optical tweezers focused to the size of a lattice spacing. We discuss three different two-qubit gates based on local collisional interactions. The gates between arbitrary qubits...... quantum computing....

6. Probing the Casimir force with optical tweezers

Ether, D S; Umrath, S; Martinez, D; Ayala, Y; Pontes, B; Araújo, G R de S; Frases, S; Ingold, G -L; Rosa, F S S; Viana, N B; Nussenzveig, H M; Neto, P A Maia

2015-01-01

We propose to use optical tweezers to probe the Casimir interaction between microspheres inside a liquid medium for geometric aspect ratios far beyond the validity of the widely employed proximity force approximation. This setup has the potential for revealing unprecedented features associated to the non-trivial role of the spherical curvatures. For a proof of concept, we measure femtonewton double layer forces between polystyrene microspheres at distances above $400$ nm by employing very soft optical tweezers, with stiffness of the order of fractions of a fN/nm. As a future application, we propose to tune the Casimir interaction between a metallic and a polystyrene microsphere in saline solution from attraction to repulsion by varying the salt concentration. With those materials, the screened Casimir interaction may have a larger magnitude than the unscreened one. This line of investigation has the potential for bringing together different fields including classical and quantum optics, statistical physics an...

7. Soft magnetic tweezers: A proof of principle

Mosconi, Francesco; Allemand, Jean François; Croquette, Vincent

2011-03-01

We present here the principle of soft magnetic tweezers which improve the traditional magnetic tweezers allowing the simultaneous application and measurement of an arbitrary torque to a deoxyribonucleic acid (DNA) molecule. They take advantage of a nonlinear coupling regime that appears when a fast rotating magnetic field is applied to a superparamagnetic bead immersed in a viscous fluid. In this work, we present the development of the technique and we compare it with other techniques capable of measuring the torque applied to the DNA molecule. In this proof of principle, we use standard electromagnets to achieve our experiments. Despite technical difficulties related to the present implementation of these electromagnets, the agreement of measurements with previous experiments is remarkable. Finally, we propose a simple way to modify the experimental design of electromagnets that should bring the performances of the device to a competitive level.

8. Electrodes for Microfluidic Integrated Optoelectronic Tweezers

Kuo-Wei Huang; Sabbir Sattar; Zhong, Jiang F.; Cheng-Hsu Chou; Hsiung-Kuang Tsai; Pei-Yu Chiou

2011-01-01

We report on two types of electrodes that enable the integration of optoelectronic tweezers (OETs) with multilayer poly(dimethylsilane)- (PDMS-) based microfluidic devices. Both types of electrodes, Au-mesh and single-walled carbon nanotube- (SWNT-) embedded PDMS thin film, are optically transparent, electrically conductive, and can be mechanically deformed and provide interfaces to form strong covalent bonding between an OET device and PDMS through standard oxygen plasma treatment. Au-mesh ...

9. A Tweezer for Chimeras in Small Networks

Omelchenko, Iryna; Omel'chenko, Oleh E.; Zakharova, Anna; Wolfrum, Matthias; Schoell, Eckehard

2015-01-01

We propose a control scheme which can stabilize and fix the position of chimera states in small networks. Chimeras consist of coexisting domains of spatially coherent and incoherent dynamics in systems of nonlocally coupled identical oscillators. Chimera states are generally difficult to observe in small networks due to their short lifetime and erratic drifting of the spatial position of the incoherent domain. The control scheme, like a tweezer, might be useful in experiments, where usually o...

10. Probing the Casimir force with optical tweezers

Ether Jr, D. S.; Pires, L. B.; Umrath, S.; Martinez, D; Ayala, Y.; Pontes, B.; Araújo, G. R. de S.; Frases, S.; Ingold, G. -L.; Rosa, F. S. S.; Viana, N. B.; Nussenzveig, H. M.; Neto, P A Maia

2015-01-01

We propose to use optical tweezers to probe the Casimir interaction between microspheres inside a liquid medium for geometric aspect ratios far beyond the validity of the widely employed proximity force approximation. This setup has the potential for revealing unprecedented features associated to the non-trivial role of the spherical curvatures. For a proof of concept, we measure femtonewton double layer forces between polystyrene microspheres at distances above $400$ nm by employing very sof...

11. Tweezers for Chimeras in Small Networks

Omelchenko, Iryna; Omel'chenko, Oleh E.; Zakharova, Anna; Wolfrum, Matthias; Schöll, Eckehard

2016-03-01

We propose a control scheme which can stabilize and fix the position of chimera states in small networks. Chimeras consist of coexisting domains of spatially coherent and incoherent dynamics in systems of nonlocally coupled identical oscillators. Chimera states are generally difficult to observe in small networks due to their short lifetime and erratic drifting of the spatial position of the incoherent domain. The control scheme, like a tweezer, might be useful in experiments, where usually only small networks can be realized.

12. Probing the Casimir force with optical tweezers

Ether, D. S., Jr.; Pires, L. B.; Umrath, S.; Martinez, D.; Ayala, Y.; Pontes, B.; Araújo, G. R. de S.; Frases, S.; Ingold, G.-L.; Rosa, F. S. S.; Viana, N. B.; Nussenzveig, H. M.; Neto, P. A. Maia

2015-11-01

We propose to use optical tweezers to probe the Casimir interaction between microspheres inside a liquid medium for geometric aspect ratios far beyond the validity of the widely employed proximity force approximation. This setup has the potential for revealing unprecedented features associated to the non-trivial role of the spherical curvatures. For a proof of concept, we measure femtonewton double-layer forces between polystyrene microspheres at distances above 400 nm by employing very soft optical tweezers, with stiffness of the order of fractions of a fN/nm. As a future application, we propose to tune the Casimir interaction between a metallic and a polystyrene microsphere in saline solution from attraction to repulsion by varying the salt concentration. With those materials, the screened Casimir interaction may have a larger magnitude than the unscreened one. This line of investigation has the potential for bringing together different fields including classical and quantum optics, statistical physics and colloid science, while paving the way for novel quantitative applications of optical tweezers in cell and molecular biology.

13. Displacement and Force Measurements with Quadrant Photodetector in Optical Tweezers

郭红莲; 刘春香; 李兆霖; 段建发; 韩学海; 程丙英; 张道中

2003-01-01

A technique of displacement and force measurements with a photodiode quadrant detector in an optical tweezers system is presented. The stiffness of optical trap is calibrated and the leukemia cell membrane tension is measured.The results show that the optical tweezers combined with the quadrant detector is a very useful tool for detecting the displacement and force with a millisecond-order response.

14. Power spectrum analysis for optical tweezers

Berg-Sørensen, K.; Flyvbjerg, H.

2004-01-01

Lorentzian provides. This is achieved using old and new theory for Brownian motion in an incompressible fluid, and new results for a popular photodetection system. The trap and photodetection system are then calibrated simultaneously in a manner that makes optical tweezers a tool of precision for force......The force exerted by an optical trap on a dielectric bead in a fluid is often found by fitting a Lorentzian to the power spectrum of Brownian motion of the bead in the trap. We present explicit functions of the experimental power spectrum that give the values of the parameters fitted, including...

15. Magnetic nanowire-enhanced optomagnetic tweezers

Mehta, Karan K.; Wu, Ting-Hsiang; Chiou, Eric P. Y.

2008-12-01

We report an optomagnetic tweezers technique that utilizes the highly localized magnetic field gradients induced near the tip of a magnetic nanowire to provide strong trapping forces on magnetic nanoparticles with high spatial resolution. Integral to our approach is a method to trap, translate, and rotate a single magnetic nanowire in three dimensions. Our simulation predicts that forces in the range of 100 pN can be generated on 200 nm magnetic particles within 0.2 μm from a 200 nm diameter nickel nanowire. Such forces are relevant in many biological processes, suggesting that this approach will be of value in biophysical studies.

16. Optical tweezers calibration with Bayesian inference

Türkcan, Silvan; Richly, Maximilian U.; Le Gall, Antoine; Fiszman, Nicolas; Masson, Jean-Baptiste; Westbrook, Nathalie; Perronet, Karen; Alexandrou, Antigoni

2014-09-01

We present a new method for calibrating an optical-tweezer setup that is based on Bayesian inference1. This method employs an algorithm previously used to analyze the confined trajectories of receptors within lipid rafts2,3. The main advantages of this method are that it does not require input parameters and is insensitive to systematic errors like the drift of the setup. Additionally, it exploits a much larger amount of the information stored in the recorded bead trajectory than standard calibration approaches. The additional information can be used to detect deviations from the perfect harmonic potential or detect environmental influences on the bead. The algorithm infers the diffusion coefficient and the potential felt by a trapped bead, and only requires the bead trajectory as input. We demonstrate that this method outperforms the equipartition method and the power-spectrum method in input information required (bead radius and trajectory length) and in output accuracy. Furthermore, by inferring a higher order potential our method can reveal deviations from the assumed second-order potential. More generally, this method can also be used for magnetic-tweezer calibration.

17. Plasmon enhanced optical tweezers with gold-coated black silicon

Kotsifaki, Domna G; Lagoudakis, Pavlos G

2016-01-01

Plasmonic optical tweezers are a ubiquitous tool for the precise manipulation of nanoparticles and biomolecules at low photon flux, while femtosecond-laser optical tweezers can probe the nonlinear optical properties of the trapped species with applications in biological diagnostics. In order to adopt plasmonic optical tweezers in real-world applications, it is essential to develop large-scale fabrication processes without compromising the trapping efficiency. Here, we develop a novel platform for continuous wave (CW) and femtosecond plasmonic optical tweezers, based on gold-coated black silicon. In contrast with traditional lithographic methods, the fabrication method relies on simple, single-step, maskless tabletop laser processing of silicon in water that facilitates scalability. Gold-coated black silicon supports repeatable trapping efficiencies comparable to the highest ones reported to date. From a more fundamental aspect, a plasmon-mediated efficiency enhancement is a resonant effect, and therefore, dep...

18. Computational toolbox for optical tweezers in geometrical optics

Callegari, Agnese; Gököz, A Burak; Volpe, Giovanni

2014-01-01

Optical tweezers have found widespread application in many fields, from physics to biology. Here, we explain in detail how optical forces and torques can be described within the geometrical optics approximation and we show that this approximation provides reliable results in agreement with experiments for particles whose characteristic dimensions are larger than the wavelength of the trapping light. Furthermore, we provide an object-oriented software package implemented in MatLab for the calculation of optical forces and torques in the geometrical optics regime: \\texttt{OTGO - Optical Tweezers in Geometrical Optics}. We provide all source codes for \\texttt{OTGO} as well as the documentation and code examples -- e.g., standard optical tweezers, optical tweezers with elongated particle, windmill effect, Kramers transitions between two optical traps -- necessary to enable users to effectively employ it in their research and teaching.

19. Local electric field measurements by optical tweezers

G. Pesce

2011-09-01

Full Text Available We report a new technique to measure direction and amplitude of electric fields generated by microelectrodes embedded in polar liquid environment, as often used in microfluidic devices. The method is based on optical tweezers which act as sensitive force transducer while a trapped charged microsphere behaves as a probe. When an electric field is applied the particles moves from its equilibrium position and finishes in a new equilibrium position where electric and optical forces are balanced. A trapped bead is moved to explore the electric field in a wide region around the microelectrodes. In such way maps of electric fields with high spatial resolution can be reconstructed even for complex electrode geometries where numerical simulation approaches can fail. Experimental results are compared with calculations based on finite element analysis simulation.

20. Single Bessel tractor-beam tweezers

Mitri, F G

2014-01-01

The tractor behavior of a zero-order Bessel acoustic beam acting on a fluid sphere, and emanating from a finite circular aperture (as opposed to waves of infinite extent) is demonstrated theoretically. Conditions for an attractive force acting in opposite direction of the radiating waves, determined by the choice of the beam's half-cone angle, the size of the radiator, and its distance from a fluid sphere, are established and discussed. Numerical predictions for the radiation force function, which is the radiation force per unit energy density and cross-sectional surface, are provided using a partial-wave expansion method stemming from the acoustic scattering. The results suggest a simple and reliable analysis for the design of Bessel beam acoustical tweezers and tractor beam devices.

1. Optical tweezers for studying taxis in parasites

In this work we present a methodology to measure force strengths and directions of living parasites with an optical tweezers setup. These measurements were used to study the parasites chemotaxis in real time. We observed behavior and measured the force of: (i) Leishmania amazonensis in the presence of two glucose gradients; (ii) Trypanosoma cruzi in the vicinity of the digestive system walls, and (iii) Trypanosoma rangeli in the vicinity of salivary glands as a function of distance. Our results clearly show a chemotactic behavior in every case. This methodology can be used to study any type of taxis, such as chemotaxis, osmotaxis, thermotaxis, phototaxis, of any kind of living microorganisms. These studies can help us to understand the microorganism sensory systems and their response function to these gradients

2. Tunable optical tweezers for wavelength-dependent measurements

Hester, Brooke; Campbell, Gretchen K.; López-Mariscal, Carlos; Filgueira, Carly Levin; Huschka, Ryan; Halas, Naomi J.; Helmerson, Kristian

2012-01-01

Optical trapping forces depend on the difference between the trap wavelength and the extinction resonances of trapped particles. This leads to a wavelength-dependent trapping force, which should allow for the optimization of optical tweezers systems, simply by choosing the best trapping wavelength for a given application. Here we present an optical tweezer system with wavelength tunability, for the study of resonance effects. With this system, the optical trap stiffness is measured for single...

3. Miniaturized Optical Tweezers Through Fiber-End Microfabrication

Liberale, Carlo

2014-07-30

Optical tweezers represent a powerful tool for a variety of applications both in biology and in physics, and their miniaturization and full integration is of great interest so as to reduce size (towards portable systems), and to minimize the required intervention from the operator. Optical fibers represent a natural solution to achieve this goal, and here we review the realization of single-fiber optical tweezers able to create a purely optical three-dimensional trap. © Springer International Publishing Switzerland 2015.

4. Control and Manipulation of Cold Atoms in Optical Tweezers

Muldoon, Cecilia; Brandt, Lukas; Dong, Jian; Stuart, Dustin; Brainis, Edouard; Himsworth, Matthew; Kuhn, Axel

2011-01-01

Neutral atoms trapped by laser light are amongst the most promising candidates for storing and processing information in a quantum computer or simulator. The application certainly calls for a scalable and flexible scheme for addressing and manipulating the atoms. We have now made this a reality by implementing a fast and versatile method to dynamically control the position of neutral atoms trapped in optical tweezers. The tweezers result from a spatial light modulator (SLM) controlling and sh...

5. Laser tweezers Raman spectroscopy of single cells

Chen, De

Raman scattering is an inelastic collision between the vibrating molecules inside the sample and the incident photons. During this process, energy exchange takes place between the photon and the scattering molecule. By measuring the energy change of the photon, the molecular vibration mode can be probed. The vibrational spectrum contains valuable information about the disposition of atomic nuclei and chemical bonds within a molecule, the chemical compositions and the interactions between the molecule and its surroundings. In this dissertation, laser tweezers Raman spectroscopy (LTRS) technique is applied for the analysis of biological cells and human cells at single cell level. In LTRS, an individual cell is trapped in aqueous medium with laser tweezers, and Raman scattering spectra from the trapped cell are recorded in real-time. The Raman spectra of these cells can be used to reveal the dynamical processes of cell growth, cell response to environment changes, and can be used as the finger print for the identification of a bacterial cell species. Several biophysical experiments were carried out using LTRS: (1) the dynamic germination process of individual spores of Bacillus thuringiensis was detected via Ca-DPA, a spore-specific biomarker molecule; (2) inactivation and killing of Bacillus subtilis spores by microwave irradiation and wet heat were studied at single cell level; (3) the heat shock activation process of single B. subtilis spores were analyzed, in which the reversible transition from glass-like state at low temperature to liquid-like state at high temperature in spore was revealed at the molecular level; (4) the kinetic processes of bacterial cell lysis of E. coli by lysozyme and by temperature induction of lambda phage were detected real-time; (5) the fixation and rehydration of human platelets were quantitatively evaluated and characterized with Raman spectroscopy method, which provided a rapid way to quantify the quality of freeze-dried therapeutic

6. Constructing Dual Beam Optical Tweezers for Undergraduate Biophysics Research

Daudelin, Brian; West-Coates, Devon; Del'Etoile, Jon; Grotzke, Eric; Paramanathan, Thayaparan

Optical tweezing, or trapping, is a modern physics technique which allows us to use the radiation pressure from laser beams to trap micron sized particles. Optical tweezers are commonly used in graduate level biophysics research but seldom used at the undergraduate level. Our goal is to construct a dual beam optical tweezers for future undergraduate biophysical research. Dual beam optical tweezers use two counter propagating laser beams to provide a stronger trap. In this study we discuss how the assembly of the dual beam optical tweezers is done through three main phases. The first phase was to construct a custom compressed air system to isolate the optical table from the vibrations from its surroundings so that we can measure pico-newton scale forces that are observed in biological systems. In addition, the biomaterial flow system was designed with a flow cell to trap biomolecules by combining several undergraduate semester projects. During the second phase we set up the optics to image and display the inside of the flow cell. Currently we are in the process of aligning the laser to create an effective trap and developing the software to control the data collection. This optical tweezers set up will enable us to study potential cancer drug interactions with DNA at the single molecule level and will be a powerful tool in promoting interdisciplinary research at the undergraduate level.

7. How safe is gamete micromanipulation by laser tweezers?

Koenig, Karsten; Tromberg, Bruce J.; Tadir, Yona; Berns, Michael W.

1998-04-01

Laser tweezers, used as novel sterile micromanipulation tools of living cells, are employed in laser-assisted in vitro fertilization (IVF). For example, controlled spermatozoa transport with 1064 nm tweezers to human egg cells has been performed in European clinics in cases of male infertility. The interaction of approximately 100 mW near infrared (NIR) trapping beams at MW/cm2 intensity with human gametes results in low mean less than 2 K temperature increases and less than 100 pN trapping forces. Therefore, photothermal or photomechanical induced destructive effects appear unlikely. However, the high photon flux densities may induce simultaneous absorption of two NIR photons resulting in nonlinear interactions. These nonlinear interactions imply non-resonant two-photon excitation of endogenous cellular chromophores. In the case of less than 800 nm tweezers, UV- like damage effects may occur. The destructive effect is amplified when multimode cw lasers are used as tweezer sources due to longitudinal mode-beating effects and partial mode- locking. Spermatozoa damage within seconds using 760 nm traps due to formation of unstable ps pulses in a cw Ti:Sa ring laser is demonstrated. We recommend the use of greater than or equal to 800 nm traps for optical gamete micromanipulation. To our opinion, further basic studies on the influence of nonlinear effects of laser tweezers on human gamete are necessary.

8. Design and Synthesis of Chiral Molecular Tweezers Based on Deoxycholic Acid

2001-01-01

A series of new chiral molecular tweezers have been designed and synthesized by using deoxycholic acid as spacer and aromatic amines as arms.Instead of using toxic phosgene,the triphosgene was employed in synthesis of the molecular tweezers receptors.These chiral molecular tweezers showed good enantioselectivity for D-amino acid methyl esters.

9. Traceable assembly of microparts using optical tweezers

Assembly of components with a size in the order of tens of micrometers or less is difficult because the gravitational forces become smaller than weak forces such as capillary, electrostatic and van der Waals forces. As such, the picked-up components commonly adhere to the manipulator, making the release operation troublesome, and the repeatable supply of components cannot be guaranteed because the magazining and bunkering scheme available in conventional scale assembly cannot be extended to these small objects. Moreover, there are also no effective ways known to deliver the finalized assembly externally. In this paper, we present the manipulation and assembly of microparts using optical tweezers, which by nature do not have stiction problems. Techniques allowing bunkering and finalizing the assembly for exporting are also presented. Finally, we demonstrate an exemplary microassembly formed by assembling two microparts: a movable microring and a microrod fixed on a glass substrate. We believe this traceable microassembly to be an important step forward for micro- and nano-manufacturing. (paper)

10. Low frequency dynamical stabilisation in optical tweezers

Richards, Christopher J.; Smart, Thomas J.; Jones, Philip H.; Cubero, David

2015-08-01

It is well known that a rigid pendulum with minimal friction will occupy a stable equilibrium position vertically upwards when its suspension point is oscillated at high frequency. The phenomenon of the inverted pendulum was explained by Kapitza by invoking a separation of timescales between the high frequency modulation and the much lower frequency pendulum motion, resulting in an effective potential with a minimum in the inverted position. We present here a study of a microscopic optical analogue of Kapitza's pendulum that operates in different regimes of both friction and driving frequency. The pendulum is realized using a microscopic particle held in a scanning optical tweezers and subject to a viscous drag force. The motion of the optical pendulum is recorded and analyzed by digital video microscopy and particle tracking to extract the trajectory and stable orientation of the particle. In these experiments we enter the regime of low driving frequency, where the period of driving is comparable to the characteristic relaxation time of the radial motion of the pendulum with finite stiffness. In this regime we find stabilization of the pendulum at angles other than the vertical (downwards) is possible for modulation amplitudes exceeding a threshold value where, unlike the truly high frequency case studied previously, both the threshold amplitude and equilibrium position are found to be functions of friction. Experimental results are complemented by an analytical theory for induced stability in the low frequency driving regime with friction.

11. A new real non-invasive single fiber tweezers

Zhang, Yu; Liu, Zhihai; Yang, Jun; Yuan, Libo

2012-02-01

A new real non-invasive two-core single fiber optical tweezers is proposed and fabricated by fiber grinding and polishing technology. The yeast cells trapping performance of this special designed truncated cone tip fiber probe is demonstrated and investigated. The distributions of the optical field emerging from the truncated cone fiber tip are simulated by Beam Prop Method. Both axial and transverse trapping forces are calculated by FDTD method. This new optical tweezers can realize truly non-invasive remote trapping and manipulating bio-cells.

12. Manipulation of microparticles and red blood cells using optoelectronic tweezers

R S Verma; R Dasgupta; N Kumar; S Ahlawat; A Uppal; P K Gupta

2014-02-01

We report the development of an optoelectronic tweezers set-up which works by lightinduced dielectrophoresis mechanism to manipulate microparticles. We used thermal evaporation technique for coating the organic polymer, titanium oxide phthalocyanine (TiOPc), as a photoconductive layer on ITO-coated glass slide. Compare to the conventional optical tweezers, the technique requires optical power in W range and provides a manipulation area of a few mm2. The set-up was used to manipulate the polystyrene microspheres and red blood cells (RBCs). The RBCs could be attracted or repelled by varying the frequency of the applied AC bias.

13. HoloHands: Kinect Control of Optical Tweezers

McDonald, Craig; McDougall, Craig; McGloin, David

2012-01-01

The increasing number of applications for holographic manipulation techniques has sparked the development of more accessible control interfaces. Here, we describe a holographic optical tweezers experiment that is controlled by gestures which are detected by a Microsoft Kinect. We demonstrate that this technique can be used to calibrate the tweezers using the Stokes Drag method and compare this to automated calibrations. We also show that multiple particle manipulation can be handled. This is a promising new line of research for gesture-based control that could find applications in a wide variety of experimental situations.

14. Marker-free cell discrimination by holographic optical tweezers

Schaal, F.; Warber, M.; Zwick, S.; van der Kuip, H.; Haist, T.; Osten, W.

2009-06-01

We introduce a method for marker-free cell discrimination based on optical tweezers. Cancerous, non-cancerous, and drug-treated cells could be distinguished by measuring the trapping forces using holographic optical tweezers. We present trapping force measurements on different cell lines: normal pre-B lymphocyte cells (BaF3; "normal cells"), their Bcr-Abl transformed counterparts (BaF3-p185; "cancer cells") as a model for chronic myeloid leukaemia (CML) and Imatinib treated BaF3-p185 cells. The results are compared with reference measurements obtained by a commercial flow cytometry system.

15. Optical tweezers reveal how proteins alter replication

Chaurasiya, Kathy

Single molecule force spectroscopy is a powerful method that explores the DNA interaction properties of proteins involved in a wide range of fundamental biological processes such as DNA replication, transcription, and repair. We use optical tweezers to capture and stretch a single DNA molecule in the presence of proteins that bind DNA and alter its mechanical properties. We quantitatively characterize the DNA binding mechanisms of proteins in order to provide a detailed understanding of their function. In this work, we focus on proteins involved in replication of Escherichia coli (E. coli ), endogenous eukaryotic retrotransposons Ty3 and LINE-1, and human immunodeficiency virus (HIV). DNA polymerases replicate the entire genome of the cell, and bind both double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) during DNA replication. The replicative DNA polymerase in the widely-studied model system E. coli is the DNA polymerase III subunit alpha (DNA pol III alpha). We use optical tweezers to determine that UmuD, a protein that regulates bacterial mutagenesis through its interactions with DNA polymerases, specifically disrupts alpha binding to ssDNA. This suggests that UmuD removes alpha from its ssDNA template to allow DNA repair proteins access to the damaged DNA, and to facilitate exchange of the replicative polymerase for an error-prone translesion synthesis (TLS) polymerase that inserts nucleotides opposite the lesions, so that bacterial DNA replication may proceed. This work demonstrates a biophysical mechanism by which E. coli cells tolerate DNA damage. Retroviruses and retrotransposons reproduce by copying their RNA genome into the nuclear DNA of their eukaryotic hosts. Retroelements encode proteins called nucleic acid chaperones, which rearrange nucleic acid secondary structure and are therefore required for successful replication. The chaperone activity of these proteins requires strong binding affinity for both single- and double-stranded nucleic

16. Plasmon enhanced optical tweezers with gold-coated black silicon

Kotsifaki, D. G.; Kandyla, M.; Lagoudakis, P. G.

2016-05-01

Plasmonic optical tweezers are a ubiquitous tool for the precise manipulation of nanoparticles and biomolecules at low photon flux, while femtosecond-laser optical tweezers can probe the nonlinear optical properties of the trapped species with applications in biological diagnostics. In order to adopt plasmonic optical tweezers in real-world applications, it is essential to develop large-scale fabrication processes without compromising the trapping efficiency. Here, we develop a novel platform for continuous wave (CW) and femtosecond plasmonic optical tweezers, based on gold-coated black silicon. In contrast with traditional lithographic methods, the fabrication method relies on simple, single-step, maskless tabletop laser processing of silicon in water that facilitates scalability. Gold-coated black silicon supports repeatable trapping efficiencies comparable to the highest ones reported to date. From a more fundamental aspect, a plasmon-mediated efficiency enhancement is a resonant effect, and therefore, dependent on the wavelength of the trapping beam. Surprisingly, a wavelength characterization of plasmon-enhanced trapping efficiencies has evaded the literature. Here, we exploit the repeatability of the recorded trapping efficiency, offered by the gold-coated black silicon platform, and perform a wavelength-dependent characterization of the trapping process, revealing the resonant character of the trapping efficiency maxima. Gold-coated black silicon is a promising platform for large-scale parallel trapping applications that will broaden the range of optical manipulation in nanoengineering, biology, and the study of collective biophotonic effects.

17. Plasmon enhanced optical tweezers with gold-coated black silicon.

Kotsifaki, D G; Kandyla, M; Lagoudakis, P G

2016-01-01

Plasmonic optical tweezers are a ubiquitous tool for the precise manipulation of nanoparticles and biomolecules at low photon flux, while femtosecond-laser optical tweezers can probe the nonlinear optical properties of the trapped species with applications in biological diagnostics. In order to adopt plasmonic optical tweezers in real-world applications, it is essential to develop large-scale fabrication processes without compromising the trapping efficiency. Here, we develop a novel platform for continuous wave (CW) and femtosecond plasmonic optical tweezers, based on gold-coated black silicon. In contrast with traditional lithographic methods, the fabrication method relies on simple, single-step, maskless tabletop laser processing of silicon in water that facilitates scalability. Gold-coated black silicon supports repeatable trapping efficiencies comparable to the highest ones reported to date. From a more fundamental aspect, a plasmon-mediated efficiency enhancement is a resonant effect, and therefore, dependent on the wavelength of the trapping beam. Surprisingly, a wavelength characterization of plasmon-enhanced trapping efficiencies has evaded the literature. Here, we exploit the repeatability of the recorded trapping efficiency, offered by the gold-coated black silicon platform, and perform a wavelength-dependent characterization of the trapping process, revealing the resonant character of the trapping efficiency maxima. Gold-coated black silicon is a promising platform for large-scale parallel trapping applications that will broaden the range of optical manipulation in nanoengineering, biology, and the study of collective biophotonic effects. PMID:27195446

18. Two- and three-beam interferometric optical tweezers

Casaburi, A.; Pesce, G.; Zemánek, Pavel; Sasso, A.

2005-01-01

Roč. 251, 4-6 (2005), s. 393-404. ISSN 0030-4018 R&D Projects: GA AV ČR(CZ) IAA1065203 Keywords : Optical force * Mie particle * Interferometric optical tweezers * Optical trapping Subject RIV: BH - Optics, Masers, Lasers Impact factor: 1.456, year: 2005

19. Scanning probe and optical tweezer investigations of biomolecular interactions

Rigby-Singleton, Shellie

2002-07-01

A complex array of intermolecular forces controls the interactions between and within biological molecules. The desire to empirically explore the fundamental forces has led to the development of several biophysical techniques. Of these, the atomic force microscope (AFM) and the optical tweezers have been employed throughout this thesis to monitor the intermolecular forces involved in biomolecular interactions. The AFM is a well-established force sensing technique capable of measuring biomolecular interactions at a single molecule level. However, its versatility has not been extrapolated to the investigation of a drug-enzyme complex. The energy landscape for the force induced dissociation of the DHFR-methotrexate complex was studied. Revealing an energy barrier to dissociation located {approx}0.3 nm from the bound state. Unfortunately, the AFM has a limited range of accessible loading rates and in order to profile the complete energy landscape alternative force sensing instrumentation should be considered, for example the BFP and optical tweezers. Thus, this thesis outlines the development and construction an optical trap capable of measuring intermolecular forces between biomolecules at the single molecule level. To demonstrate the force sensing abilities of the optical set up, proof of principle measurements were performed which investigate the interactions between proteins and polymer surfaces subjected to varying degrees of argon plasma treatment. Complementary data was gained from measurements performed independently by the AFM. Changes in polymer resistance to proteins as a response to changes in polymer surface chemistry were detected utilising both AFM and optical tweezers measurements. Finally, the AFM and optical tweezers were employed as ultrasensitive biosensors. Single molecule investigations of the antibody-antigen interaction between the cardiac troponin I marker and its complementary antibody, reveals the impact therapeutic concentrations of heparin

20. Single-sided lateral-field and phototransistor-based optoelectronic tweezers

Ohta, Aaron (Inventor); Chiou, Pei-Yu (Inventor); Hsu, Hsan-Yin (Inventor); Jamshidi, Arash (Inventor); Wu, Ming-Chiang (Inventor); Neale, Steven L. (Inventor)

2011-01-01

Described herein are single-sided lateral-field optoelectronic tweezers (LOET) devices which use photosensitive electrode arrays to create optically-induced dielectrophoretic forces in an electric field that is parallel to the plane of the device. In addition, phototransistor-based optoelectronic tweezers (PhOET) devices are described that allow for optoelectronic tweezers (OET) operation in high-conductivity physiological buffer and cell culture media.

1. Construction of an optical tweezer for nanometer scale rheology

A Raghu; Sharath Ananthamurthy

2005-10-01

The optical tweezer is a versatile set-up that can be employed in a wide variety of studies investigating the microscopic properties of materials. In particular, this set-up has in recent times been gainfully employed in probing rheological properties of materials that exhibit viscoelasticity. These measurements can provide data at the micro and nanometer scales, not normally accessible by rheometers that are used for measurements on bulk samples. In this work we describe a single laser beam optical tweezer set-up, which is built around an inverted open microscope. The trapped polystyrene particle bead's deviation from the trap potential minimum is monitored by laser back-scattering technique and the bead position measured by a quadrant photodiode detector. Additionally, a provision is made for video microscopic studies on dispersed beads using a CCD camera. A single particle microrheological experiment that can be performed using the set-up is described with relevant calculations.

2. Probing DNA with micro- and nanocapillaries and optical tweezers

Steinbock, L J; Otto, O; Skarstam, D R; Jahn, S; Chimerel, C; Gornall, J L; Keyser, U F, E-mail: ufk20@cam.ac.u [Cavendish Laboratory, University of Cambridge, J J Thomson Avenue, Cambridge CB3 0HE (United Kingdom)

2010-11-17

We combine for the first time optical tweezer experiments with the resistive pulse technique based on capillaries. Quartz glass capillaries are pulled into a conical shape with tip diameters as small as 27 nm. Here, we discuss the translocation of {lambda}-phage DNA which is driven by an electrophoretic force through the nanocapillary. The resulting change in ionic current indicates the folding state of single {lambda}-phage DNA molecules. Our flow cell design allows for the straightforward incorporation of optical tweezers. We show that a DNA molecule attached to an optically trapped colloid is pulled into a capillary by electrophoretic forces. The detected electrophoretic force is in good agreement with measurements in solid-state nanopores.

3. Synthesis and Characterization of Carbazole-Linked Porphyrin Tweezers.

Chang, Yi; Michelin, Clément; Bucher, Léo; Desbois, Nicolas; Gros, Claude P; Piant, Sébastien; Bolze, Frédéric; Fang, Yuanyuan; Jiang, Xiaoqin; Kadish, Karl M

2015-08-17

Herein the synthesis, spectroscopic characterization, two-photon absorption and electrochemical properties of 3,6-disubstituted carbazole tweezers is reported. A dimer resulting from a Glaser homocoupling was isolated during a Sonogashira coupling reaction between a diethynyl-carbazole spacer and a 5-bromo-triarylporphyrin and the properties of this original compound were compared with the 3,6-disubstituted carbazole bisporphyrin tweezers. The dyads reported herein present a two-photon absorption maximum at 920 nm with two-photon absorption cross-section in the 1200 GM range. Despite a strong linear absorption in the Soret region and moderate fluorescence quantum yield, they both lead to a high brightness reaching 30 000 M(-1)  cm(-1) . PMID:26177731

4. Continuous cell lysis in microfluidics through acoustic and optoelectronic tweezers

Witte, C.; Kremer, C; Cooper, J.M.; Neale, S.L.

2013-01-01

A versatile platform for efficient cell lysis using a combination of acoustic and electric fields in a microchannel is presented. Cell membrane disruption is triggered by electric fields inducing electroporation and then lysis. The principle of optoelectronic tweezers (OET) is applied to control the electric field strength and a surface acoustic wave transducer is attached to an OET chip to implement acoustic tweezing (AT). The system is characterized in terms of spatial contro...

5. Optical tweezers for the study of microbubble dynamics in ultrasound

Garbin, Valeria

2007-01-01

Optical tweezers enable for non-destructive, contact-free manipulation of ultrasound contrast agent (UCA) microbubbles, which are used in medical imaging for enhancing the echogenicity of the blood pool and to quantify organ perfusion. Understanding the dynamics of ultrasound-driven contrast agent microbubbles from a fundamental physical standpoint is a first step for exploiting their acoustical properties and to develop new diagnostic and therapeutic applications. However, ...

6. Mechanical Forces Impeding Exocytotic Surfactant Release Revealed by Optical Tweezers

Singer, Wolfgang; Frick, Manfred; Haller, Thomas; Bernet, Stefan; Ritsch-Marte, Monika; Dietl, Paul

2003-01-01

The release of surfactant from alveolar type II cells is essential to lower the surface tension in the lung and to facilitate inspiration. However, the factors controlling dispersal and diffusion of this hydrophobic material are still poorly understood. Here we report that release of surfactant from the fused vesicle, termed lamellar body (LB), resisted mechanical forces applied by optical tweezers: At constant trapping force, the probability to expand LB contents, i.e., to “pull” surfactant ...

7. Optical tweezers and manipulation of PMMA beads in various conditions

Kotsifaki, D. G.; Makropoulou, M.; Serafetinides, A. A.

2009-07-01

Laser optical trapping and micromanipulation of microparticles or cells and subcellular structures have gained remarkable interest in biomedical research and applications. Several laser sources are employed for the combination of a laser scalpel with an optical tweezers device, under microscopic control. However, although the principles and the mechanisms of pulsed laser ablation have been well described for macroscopic interventions, the microbeam operation, under microscopic guidance, necessitates further experiments and investigations. We present experimental results of controlled micro-ablation of PMMA beads of 3-8 μm diameters, trapped by laser tweezers in various media e.g. solutes of different index of refraction. An optical tweezers system, based on a continuous wave He-Ne laser emitting at 632.8 nm, was tested on beads and, despite the low power of the He-Ne laser, the optical trap was stable. Another optical system, based on a cw Nd:YAG laser emitting at 1.06 μm, was tested on microspheres too. Successful beads ablation was carried out by irradiation with multiple, or even a single nitrogen laser pulse of 7 ns pulse duration at a wavelength of 337 nm. The ablative perforation of the microspheres was estimated by controlling the laser fluence. Moreover, shape deformations of PMMA microspheres were observed. The experimentally obtained results are theoretically explained via the spatial intensity distribution based on Mie light scattering theory. Furthermore, the appearance of laser ablation holes in the back side of microspheres is explained by the ablation triggered shock waves propagation. The role of the stretching forces action is also discussed. Additionally, we report experimental results on measuring the optical trap force of PMMA beads. A powerful optical tweezers system based on a continuous wave Nd:YAG laser was used in order to estimate the trapping efficiency for several beads diameter.

8. Optical Fiber Tweezers Fabricated by Guided Wave Photo-Polymerization

Rita S. Rodrigues Ribeiro; Raquel Queirós; Olivier Soppera; Ariel Guerreiro; Jorge, Pedro A. S.

2015-01-01

In this work the use of guided wave photo-polymerization for the fabrication of novel polymeric micro tips for optical trapping is demonstrated. It is shown that the selective excitation of linear polarized modes, during the fabrication process, has a direct impact on the shape of the resulting micro structures. Tips are fabricated with modes LP02 and LP21 and their shapes and output intensity distribution are compared. The application of the micro structures as optical tweezers is demonstrat...

9. Cluster formation in ferrofluids induced by holographic optical tweezers.

Masajada, Jan; Bacia, Marcin; Drobczyński, Sławomir

2013-10-01

Holographic optical tweezers were used to show the interaction between a strongly focused laser beam and magnetic nanoparticles in ferrofluid. When the light intensity was high enough, magnetic nanoparticles were removed from the beam center and formed a dark ring. The same behavior was observed when focusing vortex or Bessel beams. The interactions between two or more separated rings of magnetic nanoparticles created by independent optical traps were also observed. PMID:24081086

10. Dynamic array generation and pattern formation for optical tweezers

2000-01-01

The generalised phase contrast approach is used for the generation of optical arrays of arbitrary beam shape, suitable for applications in optical tweezers for the manipulation of biological specimens. This approach offers numerous advantages over current techniques involving the use of computer-......-generated holograms or diffractive optical elements. We demonstrate a low-loss system for generating intensity patterns suitable for the trapping and manipulation of small particles or specimens....

11. Expression of Echmr gene from Eichhornia offers multiple stress tolerance to Cd sensitive Escherichia coli Δgsh mutants.

Thapa, G; Das, D; Gunupuru, L R

2016-09-01

The detoxification of heavy metals frequently involves conjugation to glutathione prior to compartmentalization and eflux in higher plants. We have expressed a heavy metal stress responsive (Echmr) gene from water hyacinth, which conferred tolerance to Cd sensitive Escherichia coli Δgsh mutants against heavy metals and abiotic stresses. The recombinant E. coli Δgsh mutant cells showed better growth recovery and survival than control cells under Cd (200 μM), Pb(200 μM), heat shock (50 °C), cold stress at 4 °C for 4 h, and UV-B (20 min) exposure. The enhanced expression of Echmr gene revealed by northern analysis during above stresses further advocates its role in multi-stress tolerance. Heterologous expression of EcHMR from Eichhornia rescued Cd(2+) sensitive E. coli mutants from Cd(2+) toxicity and induced better recovery post abiotic stresses. This may suggests a possible role of Echmr in Cd(II) and desiccation tolerance in plants for enhanced stress response. PMID:27457806

12. Linear microrheology with optical tweezers of living cells 'is not an option'!

Tassieri, Manlio

2015-08-01

Optical tweezers have been successfully adopted as exceptionally sensitive transducers for microrheology studies of complex fluids. Despite the general trend, in this article I explain why a similar approach should not be adopted for microrheology studies of living cells. This conclusion is acheived on the basis of statistical mechanics principles that indicate the unsuitability of optical tweezers for such purpose. PMID:26100967

13. Linear microrheology with optical tweezers of living cells 'is not an option'!

Tassieri, Manlio

2015-01-01

Optical tweezers have been successfully adopted as exceptionally sensitive transducers for microrheology studies of complex fluids. Despite the general trend, in this article I explain why a similar approach should not be adopted for microrheology studies of living cells. This conclusion is reached on the basis of statistical mechanics principles that indicate the unsuitability of optical tweezers for such purpose.

14. Fiber-pigtailed optical tweezer for single-atom trapping and single-photon generation

Garcia, Sébastien; Hohmann, Leander; Reichel, Jakob; Long, Romain

2013-01-01

We demonstrate a miniature, fiber-coupled optical tweezer to trap a single atom. The same fiber is used to trap a single atom and to read out its fluorescence. To obtain a low background level, the tweezer light is chopped, and we measure the influence of the chopping frequency on the atom's lifetime. We use the single atom as a single-photon source at 780 nm and measure the second-order correlation function of the emitted photons. Because of its miniature, robust, fiber-pigtailed design, this tweezer can be implemented in a broad range of experiments where single atoms are used as a resource.

15. Optical sorting of particles by dual-channel line optical tweezers

A novel configuration of dual-channel line optical tweezers with a ‘Y’ shape is constructed for sorting of particles within a microfluidic chip. When yeast cells with different size pass the intersection of the specially designed line optical tweezers, they are separated and transported to different channels due to a difference in the forces exerted by the line tweezers that depends on the size of the cells. The influences of some experimental conditions, such as laser power and flow velocity, on the sorting efficiency are discussed. (paper)

16. TweezPal - Optical tweezers analysis and calibration software

Osterman, Natan

2010-11-01

Optical tweezers, a powerful tool for optical trapping, micromanipulation and force transduction, have in recent years become a standard technique commonly used in many research laboratories and university courses. Knowledge about the optical force acting on a trapped object can be gained only after a calibration procedure which has to be performed (by an expert) for each type of trapped objects. In this paper we present TweezPal, a user-friendly, standalone Windows software tool for optical tweezers analysis and calibration. Using TweezPal, the procedure can be performed in a matter of minutes even by non-expert users. The calibration is based on the Brownian motion of a particle trapped in a stationary optical trap, which is being monitored using video or photodiode detection. The particle trajectory is imported into the software which instantly calculates position histogram, trapping potential, stiffness and anisotropy. Program summaryProgram title: TweezPal Catalogue identifier: AEGR_v1_0 Program summary URL:http://cpc.cs.qub.ac.uk/summaries/AEGR_v1_0.html Program obtainable from: CPC Program Library, Queen's University, Belfast, N. Ireland Licensing provisions: Standard CPC licence, http://cpc.cs.qub.ac.uk/licence/licence.html No. of lines in distributed program, including test data, etc.: 44 891 No. of bytes in distributed program, including test data, etc.: 792 653 Distribution format: tar.gz Programming language: Borland Delphi Computer: Any PC running Microsoft Windows Operating system: Windows 95, 98, 2000, XP, Vista, 7 RAM: 12 Mbytes Classification: 3, 4.14, 18, 23 Nature of problem: Quick, robust and user-friendly calibration and analysis of optical tweezers. The optical trap is calibrated from the trajectory of a trapped particle undergoing Brownian motion in a stationary optical trap (input data) using two methods. Solution method: Elimination of the experimental drift in position data. Direct calculation of the trap stiffness from the positional

17. Efficient loading of a single neutral atom into an optical microscopic tweezer

何军; 刘贝; 刁文婷; 王杰英; 靳刚; 王军民

2015-01-01

A single atom in a magneto–optical trap (MOT) with trap size (hundreds of micrometers) can be transferred into an optical microscopic tweezer with a probability of∼100%. The ability to transfer a single atom into two traps back and forth allows us to study the loading process. The loading probability is found to be insensitive to the geometric overlap of the MOT and the tweezer. It is therefore possible to perform simultaneously loading of a single atom into all sites of the tweezer array for many qubits. In particular, we present a simulation of the one-dimensional and two-dimensional arrays of an optical microscopic tweezer. We find the same qualitative behavior for all of the trap parameters.

18. Construction of force measuring optical tweezers instrumentation and investigations of biophysical properties of bacterial adhesion organelles

2015-01-01

Optical tweezers are a technique in which microscopic-sized particles, including living cells and bacteria, can be non-intrusively trapped with high accuracy solely using focused light. The technique has therefore become a powerful tool in the field of biophysics. Optical tweezers thereby provide outstanding manipulation possibilities of cells as well as semi-transparent materials, both non-invasively and non-destructively, in biological systems. In addition, optical tweezers can measure minute forces (< 10-12 N), probe molecular interactions and their energy landscapes, and apply both static and dynamic forces in biological systems in a controlled manner. The assessment of intermolecular forces with force measuring optical tweezers, and thereby the biomechanical structure of biological objects, has therefore considerably facilitated our understanding of interactions and structures of biological systems. Adhesive bacterial organelles, so called pili, mediate adhesion to host cells and are therefore crucial...

19. A study of multi-trapping of tapered-tip single fiber optical tweezers

We develop a pair of tapered-tip single fiber optical tweezers, and study its multi-trapping characteristic. The finite difference time domain method is employed to simulate the trapping force characteristic of this pair of single fiber optical tweezers, and the results show that the number of trapped particles depends on the refractive index and the size of the particles. The trapping force of this pair of tapered-tip single fiber optical tweezers is calibrated by the experimental method, and the experimental results are consistent with the theoretical calculation results. The multi-trapping capability realized by the tapered-tip single fiber optical tweezers will be practical and useful for applications in biomedical research fields. (interdisciplinary physics and related areas of science and technology)

20. Application of BP neural networks in non-linearity correction of optical tweezers

Ziqiang WANG; Yinmei LI; Liren LOU; Henghua WEI; Zhong WANG

2008-01-01

The back-propagation (BP) neural network is proposed to correct nonlinearity and optimize the force measurement and calibration of an optical tweezer sys-tem. Considering the low convergence rate of the BP algo-rithm, the Levenberg-Marquardt (LM) algorithm is used to improve the BP network. The proposed method is experimentally studied for force calibration in a typical optical tweezer system using hydromechanics. The result shows that with the nonlinear correction using BP net-works, the range of force measurement of an optical tweezer system is enlarged by 30% and the precision is also improved compared with the polynomial fitting method. It is demonstrated that nonlinear correction by the neural network method effectively improves the per-formance of optical tweezers without adding or changing the measuring system.

1. Optical nanofiber integrated into an optical tweezers for particle probing and manipulation

Frawley, Mary C; Truong, Viet Giang; Chormaic, Sile Nic

2014-01-01

We present an integrated platform for particle manipulation consisting of a combined optical nanofiber and optical tweezers system. Individual silica microspheres were introduced to the nanofiber at arbitrary points using the optical tweezers, thereby producing pronounced dips in the fiber transmission. We show that such consistent and reversible transmission modulations depend on both particle and fiber diameter, and may be used as a reference point for in-situ nanofiber or particle size calibration. Particle arrays can be released from the optical tweezers onto the nanofiber and are propelled along the fiber length via guided light. We also demonstrate how the optical tweezers can be used to create a "particle jet" to feed a supply of microspheres to the nanofiber surface, forming a particle conveyor belt. This integrated optical platform provides a method for selective evanescent field manipulation of micron-sized particles and may facilitate studies of optical binding and light-particle interaction dynami...

2. Extending a release-and-recapture scheme to single atom optical tweezer for effective temperature evaluation

By recording the fluorescence fraction of the cold atoms remaining in the magneto-optical trap (MOT) as a function of the release time, the release-and-recapture (R and R) method is utilized to evaluate the effective temperature of the cold atomic ensemble. We prepare a single atom in a large-magnetic-gradient MOT and then transfer the trapped single atom into a 1064-nm microscopic optical tweezer. The energy of the single atom trapped in the tweezer is further reduced by polarization gradient cooling (PGC) and the effective temperature is evaluated by extending the R and R technique to a single atom tweezer. The typical effective temperature of a single atom in the tweezer is improved from about 105 μK to about 17 μK by applying the optimum PGC phase. (atomic and molecular physics)

3. Design and construction of an optical compact and affordable tweezers

The following paper presents a new design allowing a reduction on the amount of required optical elements for the construction of the optical tweezers, which results in a compact and affordable system. The latter is composed by a 40 mW Nd: YAG (532 nm) laser and commercially available optics and mounts. A virtual instrument, developed using Lab View 8.0, controls both a XYZ table and a web camera for visualizing. The quality of the laser beam at the end of optical system was characterized using the knife-edge method, resulting in a Gaussian shaped (TEM00) intensity profile. It is presented as well the necessary information to align and calibrate each optical element. (Author)

4. Microrheology of concentrated DNA solutions using optical tweezers

Arun S Rajkumar; B M Jaffar Ali

2008-06-01

Semiflexible biopolymers play a vital role in shaping cellular structure and rigidity. In this work, we report the determination of microrheological properties of concentrated, double-stranded calf thymus DNA (CT-DNA) solutions using passive, laser-scattering based particle-tracking methodology. From power spectral analysis, we obtain dynamic shear moduli of the polymer solutions stretching over three decades of frequency (100–103 Hz) and over concentration ranges spanning from very dilute to concentrated regime. We also study the effects of altered ionic strength and denaturation on the shear modulus. Our results indicate that (CT-DNA) exhibits predominantly elastic behaviour in the concentration range we probed. From the measurements of the plateau shear modulus, p, we conclude that DNA generally behaves like a semiflexible polymer in a good solvent even at low ionic strength. We have thus demonstrated application of passive microrheological method using optical tweezers to DNA solutions. Further extensions of the technique and its applications are discussed.

5. Dynamic properties of bacterial pili measured by optical tweezers

Fallman, Erik; Schedin, Staffan; Jass, Jana; Uhlin, Bernt Eric; Axner, Ove

2014-01-01

The ability of uropathogenic Escherichia coli (UPEC) to cause urinary tract infections is dependent on their ability to colonize the uroepithelium. Infecting bacteria ascend the urethra to the bladder and then kidneys by attaching to the uroepithelial cells via the differential expression of adhesins. P pili are associated with pyelonephritis, the more severe infection of the kidneys. In order to find means to treat pyelonephritis, it is therefore of interest to investigate the properties P pili. The mechanical behavior of individual P pili of uropathogenic Escherichia coli has recently been investigated using optical tweezers. P pili, whose main part constitutes the PapA rod, composed of ~1000 PapA subunits in a helical arrangement, are distributed over the bacterial surface and mediate adhesion to host cells. We have earlier studied P pili regarding its stretching/elongation properties where we have found and characterized three different elongation regions, of which one constitute an unfolding of the quate...

6. iTweezers: optical micromanipulation controlled by an Apple iPad

Bowman, R. W.; Gibson, G.; Carberry, D.; Picco, L.; Miles, M.; Padgett, M. J.

2011-04-01

The 3D interactive manipulation of multiple particles with holographic optical tweezers is often hampered by the control system. We use a multi-touch interface implemented on an Apple iPad to overcome many of the limitations of mouse-based control, and demonstrate an elegant and intuitive interface to multi-particle manipulation. This interface connects to the tweezers system hardware over a wireless network, allowing it to function as a remote monitor and control device.

7. Interrogating Biology with Force: Single Molecule High-Resolution Measurements with Optical Tweezers

Capitanio, Marco; Pavone, Francesco S.

2013-01-01

Single molecule force spectroscopy methods, such as optical and magnetic tweezers and atomic force microscopy, have opened up the possibility to study biological processes regulated by force, dynamics of structural conformations of proteins and nucleic acids, and load-dependent kinetics of molecular interactions. Among the various tools available today, optical tweezers have recently seen great progress in terms of spatial resolution, which now allows the measurement of atomic-scale conformat...

8. Measurement of interaction forces between red blood cells in aggregates by optical tweezers

Maklygin, A Yu; Priezzhev, A V; Karmenian, A; Nikitin, Sergei Yu; Obolenskii, I S; Lugovtsov, Andrei E; Kisun Li

2012-06-30

We have fabricated double-beam optical tweezers and demonstrated the possibility of their use for measuring the interaction forces between red blood cells (erythrocytes). It has been established experimentally that prolonged trapping of red blood cells in a tightly focused laser beam does not cause any visible changes in their shape or size. We have measured the interaction between red blood cells in the aggregate, deformed by optical tweezers.

9. Fiber optical tweezers for microscale and nanoscale particle manipulation and force sensing

Liu, Yuxiang

2011-12-01

Optical tweezers have been an important tool in biology and physics for studying single molecules and colloidal systems. Most of current optical tweezers are built with microscope objectives, which are: i) expensive, ii) bulky and hard to integrate, iii) sensitive to environmental fluctuations, iv) limited in terms of working distances from the substrate, and v) rigid with the requirements on the substrate (transparent substrate made with glass and with a fixed thickness). These limitations of objective-based optical tweezers prevent them from being miniaturized. Fiber optical tweezers can provide a solution for cost reduction and miniaturization, and these optical tweezers can be potentially used in microfluidic systems. However, the existing fiber optical tweezers have the following limitations: i) low trapping efficiency due to weakly focused beams, ii) lack of the ability to control the positions of multiple particles simultaneously, and iii) limited functionalities. The overall objective of this dissertation work is to further the fundamental understanding of fiber optical tweezers through experimental study and modeling, and to develop novel fiber optical tweezers systems to enhance the capability and functionalities of fiber optical tweezers as microscale and nanoscale manipulators/sensors. The contributions of this dissertation work are summarized as follows. i) An enhanced understanding of the inclined dual-fiber optical tweezers (DFOTs) system has been achieved. Stable three dimensional (3D) optical trapping of a single micron-sized particle has been experimentally demonstrated. This is the first time that the trapping efficiency has been calibrated and the stiffness of the trap has been obtained in the experiments, which has been carried out by using two methods: the drag force method and power spectrum analysis. Such calibration enables the system to be used as a picoNewton-level force sensor in addition to a particle manipulator. The influence of

10. Static and dynamic response of carbon nano tube-based nano-tweezers

In this paper static and dynamic responses of a nano-tweezer composed of two carbon nano-tube arms are investigated. Taking into account a continuum model and considering the electrostatic actuation as well as the presence of the van der Waals forces, the static nonlinear equations are solved by a step by step linearization and Galerkin projection method. Simulating the closing dynamics of the nano-tweezer, the specified effective diameters of the nano tubes compared to existing experimental data. Then by imposing a step DC voltage and taking into account the inertia effects, the dynamic responses and pull-in conditions of the nano-tweezer are studied. In the static and dynamic analysis, the effects of various parameters such as initial gap, length and diameter of the nano tubes on the pull-in conditions are investigated. Also the effect of damping and asymmetric stiffness of the arms on the pull-in voltages of the nano-tweezer is reported. Comparison of the results with the published experimental data shows that the use of continuum model and employing the Galerkin based step by step linearization method could effectively simulate the response of nano-tweezers.

11. Multiplexed single-molecule measurements with magnetic tweezers

We present a method for performing multiple single-molecule manipulation experiments in parallel with magnetic tweezers. We use a microscope with a low magnification, and thus a wide field of view, to visualize multiple DNA-tethered paramagnetic beads and apply an optimized image analysis routine to track the three-dimensional position of each bead simultaneously in real time. Force is applied to each bead using an externally applied magnetic field. Since variations in the field parameters are negligible across the field of view, nearly identical manipulation of all visible beads is possible. However, we find that the error in the position measurement is inversely proportional to the microscope's magnification. To mitigate the increased error caused by demagnification, we have developed a strategy based on tracking multiple fixed beads. Our system is capable of simultaneously manipulating and tracking up to 34 DNA-tethered beads at 60 Hz with ∼1.5 nm resolution and with ∼10% variation in applied force.

12. Accurate measurement of microscopic forces and torques using optical tweezers

Andrew Forbes

2011-09-01

Full Text Available It is now well known that matter may be trapped by optical fields with high intensity gradients. Once trapped, it is then possible to manipulate microscopic particles using such optical fields, in so-called optical tweezers. Such optical trapping and tweezing systems have found widespread application across diverse fields in science, from applied biology to fundamental physics. In this article we outline the design and construction of an optical trapping and tweezing system, and show how the resulting interaction of the laser light with microscopic particles may be understood in terms of the transfer of linear and angular momentum of light. We demonstrate experimentally the use of our optical tweezing configuration for the measurement of microscopic forces and torques. In particular, we make use of digital holography to create so-called vortex laser beams, capable of transferring orbital angular momentum to particles. The use of such novel laser beams in an optical trapping and tweezing set-up allows for the control of biological species at the single-cell level.

13. Dispersive light-matter interaction in programmable optical tweezers

Sawyer, Bianca J.; Horvath, Milena S. J.; Deb, Amita B.; Kjørgaard, Niels

2015-08-01

We have developed a robust interrogation system using frequency modulation spectroscopy to measure the quantum state-dependent phase shift incurred on an off-resonant optical probe when transmitted by an atomic medium. Recently, our focus has been on extending this technique for the detection of Feshbach resonances in 87Rb atoms. Feshbach resonance is a mechanism which allows the atomic interaction strength to be precisely tuned via an external magnetic field. To access a Feshbach resonance atoms must be independently prepared in certain internal states, during which we utilize programmable optical tweezers to perform precise spatial micro-manipulation of the ensemble in laser "test-tubes." We use our dispersive probing system to identify the resonant magnetic field value in a sample with a dense "ball" geometry. An important design consideration for such a probing scheme is the three-dimensional mode-matching at the interface between light and the atomic sample when coupled by the dispersive interaction. We discuss challenges which dealing with this new geometry compared to the previously used prolate geometry, and consider the possibility of dipole-dipole interactions in our sample leading to cooperative light scattering processes.

14. Calibration of femtosecond optical tweezers as a sensitive thermometer

Mondal, Dipankar; Goswami, Debabrata

2015-08-01

We present cumulative perturbation effects of femtosecond laser pulses on an optical tweezer. Our experiments involve a dual wavelength high repetition rate femtosecond laser, one at the non-heating wavelength of 780 nm while the other at 1560 nm to cause heating in the trapped volume under low power (100-800 μW) conditions. The 1560 nm high repetition rate laser acts as a resonant excitation source for the vibrational combination band of the hydroxyl group (OH) of water, which helps create the local heating effortlessly within the trapping volume. With such an experimental system, we are the first to observe direct effect of temperature on the corner frequency deduced from power spectrum. We can, thus, control and measure temperature precisely at the optical trap. This observation has lead us to calculate viscosity as well as temperature in the vicinity of the trapping zone. These experimental results also support the well-known fact that the nature of Brownian motion is the response of the optically trapped bead from the temperature change of surroundings. Temperature rise near the trapping zone can significantly change the viscosity of the medium. However, we notice that though the temperature and viscosity are changing as per our corner frequency calculations, the trap stiffness remains the same throughout our experiments within the temperature range of about 20 K.

15. Hong-Ou-Mandel atom interferometry in tunnel-coupled optical tweezers

Lester, Brian; Kaufman, Adam; Reynolds, Collin; Wall, Michael; Foss-Feig, Michael; Hazzard, Kaden; Rey, Ana Maria; Regal, Cindy

2014-05-01

We present recent work in which we demonstrate near-complete control over all the internal and external degrees of freedom of laser-cooled 87Rb atoms trapped in sub-micron optical tweezers. Utilizing this control for two atoms in two optical tweezers, we implement a massive-particle analog of the Hong-Ou-Mandel interferometer where atom tunneling plays the role of the photon beamsplitter. The interferometer is used to probe the effect of atomic indistinguishability on the two-atom dynamics for a variety of initial conditions. These experiments demonstrate the viability of the optical tweezer platform for bottom-up generation of low-entropy quantum systems and pave the way toward the direct observation of quantum dynamics in more complex finite-sized systems.

16. Optical disassembly of cellular clusters by tunable tug-of-war tweezers

Bezryadina, Anna; Chen, Joseph C; Chen, Zhigang

2016-01-01

Bacterial biofilms underlie many persistent infections, posing major hurdles in antibiotic treatment. Here, we design and demonstrate tug-of-war optical tweezers that can facilitate assessment of cell-cell adhesion - a key contributing factor to biofilm development, thanks to the combined actions of optical scattering and gradient forces. With a customized optical landscape distinct from that of conventional tweezers, not only can such tug-of-war tweezers stably trap and stretch a rod-shaped bacterium in the observing plane, but, more importantly, they can also impose a tunable lateral force that pulls apart cellular clusters without any tethering or mechanical movement. As a proof of principle, we examined a Sinorhizobium meliloti strain that forms robust biofilms and found that the strength of intercellular adhesion depends on the growth medium. This technique may herald new photonic tools for optical manipulation and biofilm study, as well as other biological applications.

17. Manipulation of Nanoparticles Using Dark-Field-Illumination Optical Tweezers with Compensating Spherical Aberration

ZHOU Jin-Hua; TAO Run-Zhe; HU Zhi-Bin; ZHONG Min-Cheng; WANG Zi-Qiang; CAI Jun; LI Yin-Mei

2009-01-01

Based on our previous investigation of optical tweezers with dark field illumination[Chin.Phys.Lett.25(2008)329],nanoparticles at large trap depth axe better viewed in wide field and real time for a long time,but with poor forces.Here we present the mismatched tube length to compensate for spherical aberration of an oil-immersion objective in a glass-water interface in an optical tweezers system for manipulating nanoparticles.In this way,the critical power of stable trapping particles is measured at different trap depths.It is found that trap depth is enlarged for trapping nanoparticles and trapping forces are enhanced at large trap depth.According to the measurement,70-nm particles are manipulated in three dimensions and observed clearly at large appropriate depth.This will expand applications of optical tweezers in a nanometre-scale colloidal system.

18. Near-field enhanced optical tweezers utilizing femtosecond-laser nanostructured substrates

We present experimental evidence of plasmonic-enhanced optical tweezers, of polystyrene beads in deionized water in the vicinity of metal-coated nanostructures. The optical tweezers operate with a continuous wave near-infrared laser. We employ a Cu/Au bilayer that significantly improves dissipation of heat generated by the trapping laser beam and avoid de-trapping from heat convection currents. We investigate the improvement of the optical trapping force and the effective trapping quality factor, and observe an exponential distance dependence of the trapping force from the nanostructures, indicative of evanescent plasmonic enhancement

19. Single DNA molecule grafting and manipulation using a combined atomic force microscope and an optical tweezer

Shivashankar, G. V.; Libchaber, A.

1997-12-01

In this letter, we report on spatially selecting and grafting a DNA-tethered bead to an atomic force microscope (AFM) cantilever, using an optical tweezer. To quantify this technique, we measure force versus extension of a single DNA molecule using AFM. For such studies, we have developed a micromanipulation approach by combining an AFM, an optical tweezer, and visualization setup. The ability to select a single DNA polymer and specifically graft it to a localized position on a substrate opens up new possibilities in biosensors and bioelectronic devices.

20. Substrate-dependent cell elasticity measured by optical tweezers indentation

Yousafzai, Muhammad S.; Ndoye, Fatou; Coceano, Giovanna; Niemela, Joseph; Bonin, Serena; Scoles, Giacinto; Cojoc, Dan

2016-01-01

In the last decade, cell elasticity has been widely investigated as a potential label free indicator for cellular alteration in different diseases, cancer included. Cell elasticity can be locally measured by pulling membrane tethers, stretching or indenting the cell using optical tweezers. In this paper, we propose a simple approach to perform cell indentation at pN forces by axially moving the cell against a trapped microbead. The elastic modulus is calculated using the Hertz-model. Besides the axial component, the setup also allows us to examine the lateral cell-bead interaction. This technique has been applied to measure the local elasticity of HBL-100 cells, an immortalized human cell line, originally derived from the milk of a woman with no evidence of breast cancer lesions. In addition, we have studied the influence of substrate stiffness on cell elasticity by performing experiments on cells cultured on two substrates, bare and collagen-coated, having different stiffness. The mean value of the cell elastic modulus measured during indentation was 26±9 Pa for the bare substrate, while for the collagen-coated substrate it diminished to 19±7 Pa. The same trend was obtained for the elastic modulus measured during the retraction of the cell: 23±10 Pa and 13±7 Pa, respectively. These results show the cells adapt their stiffness to that of the substrate and demonstrate the potential of this setup for low-force probing of modifications to cell mechanics induced by the surrounding environment (e.g. extracellular matrix or other cells).

1. Dynamic properties of bacterial pili measured by optical tweezers

Fallman, Erik G.; Andersson, Magnus J.; Schedin, Staffan S.; Jass, Jana; Uhlin, Bernt Eric; Axner, Ove

2004-10-01

The ability of uropathogenic Escherichia coli (UPEC) to cause urinary tract infections is dependent on their ability to colonize the uroepithelium. Infecting bacteria ascend the urethra to the bladder and then kidneys by attaching to the uroepithelial cells via the differential expression of adhesins. P pili are associated with pyelonephritis, the more severe infection of the kidneys. In order to find means to treat pyelonephritis, it is therefore of interest to investigate the properties P pili. The mechanical behavior of individual P pili of uropathogenic Escherichia coli has recently been investigated using optical tweezers. P pili, whose main part constitutes the PapA rod, composed of ~1000 PapA subunits in a helical arrangement, are distributed over the bacterial surface and mediate adhesion to host cells. We have earlier studied P pili regarding its stretching/elongation properties where we have found and characterized three different elongation regions, of which one constitute an unfolding of the quaternary (helical) structure of the PapA rod. It was shown that this unfolding takes place at an elongation independent force of 27 +/- 2 pN. We have also recently performed studies on its folding properties and shown that the unfolding/folding of the PapA rod is completely reversible. Here we present a study of the dynamical properties of the PapA rod. We show, among other things, that the unfolding force increases and that the folding force decreases with the speed of unfolding and folding respectively. Moreover, the PapA rod can be folded-unfolded a significant number of times without loosing its characteristics, a phenomenon that is believed to be important for the bacterium to keep close contact to the host tissue and consequently helps the bacterium to colonize the host tissue.

2. Combining optical tweezers and scanning probe microscopy to study DNA-protein interactions

Huisstede, Jurgen H.G.; Subramaniam, Vinod; Bennink, Martin L.

2007-01-01

We present the first results obtained with a new instrument designed and built to study DNA-protein interactions at the single molecule level. This microscope combines optical tweezers with scanning probe microscopy and allows us to locate DNA-binding proteins on a single suspended DNA molecule. A s

3. Research Advances: Nanoscale Molecular Tweezers; Cinnamon as Pesticide?; Recently Identified Dietary Sources of Antioxidants

King, Angela G.

2004-12-01

This Report from Other Journals surveys articles of interest to chemists that have been recently published in other science journals. Topics surveyed include reports that receptors have been designed to act as molecular tweezers; cinnamon has potential in the fight against mosquitoes; and high levels of antioxidants are found in some surprising foods. See Featured Molecules .

4. Holographic Raman tweezers controlled by multi-modal natural user interface

Tomori, Z.; Keša, P.; Nikorovič, M.; Kaňka, Jan; Jákl, Petr; Šerý, Mojmír; Bernatová, Silvie; Valušová, E.; Antalík, M.; Zemánek, Pavel

2016-01-01

Roč. 18, č. 1 (2016), 015602:1-9. ISSN 2040-8978 R&D Projects: GA MŠk(CZ) LO1212; GA MŠk(CZ) LD14069 Institutional support: RVO:68081731 Keywords : holographic optical tweezers * Raman microspectroscopy * human-computer interface Subject RIV: BH - Optics, Masers, Laser s Impact factor: 2.059, year: 2014

5. High Spatiotemporal-Resolution Magnetic Tweezers: Calibration and Applications for DNA Dynamics.

Dulin, David; Cui, Tao Ju; Cnossen, Jelmer; Docter, Margreet W; Lipfert, Jan; Dekker, Nynke H

2015-11-17

The observation of biological processes at the molecular scale in real time requires high spatial and temporal resolution. Magnetic tweezers are straightforward to implement, free of radiation or photodamage, and provide ample multiplexing capability, but their spatiotemporal resolution has lagged behind that of other single-molecule manipulation techniques, notably optical tweezers and AFM. Here, we present, to our knowledge, a new high-resolution magnetic tweezers apparatus. We systematically characterize the achievable spatiotemporal resolution for both incoherent and coherent light sources, different types and sizes of beads, and different types and lengths of tethered molecules. Using a bright coherent laser source for illumination and tracking at 6 kHz, we resolve 3 Å steps with a 1 s period for surface-melted beads and 5 Å steps with a 0.5 s period for double-stranded-dsDNA-tethered beads, in good agreement with a model of stochastic bead motion in the magnetic tweezers. We demonstrate how this instrument can be used to monitor the opening and closing of a DNA hairpin on millisecond timescales in real time, together with attendant changes in the hairpin dynamics upon the addition of deoxythymidine triphosphate. Our approach opens up the possibility of observing biological events at submillisecond timescales with subnanometer resolution using camera-based detection. PMID:26588570

6. High-refractive index particles in counter-propagating optical tweezers - manipulation and forces

Horst, Astrid van der

2006-01-01

With a tightly focused single laser beam, also called optical tweezers, particles of a few nanometers up to several micrometers in size can be trapped and manipulated in 3D. The size, shape and refractive index of such colloidal particles are of influence on the optical forces exerted on them in the

7. Construction and actuation of a microscopic gear assembly formed using optical tweezers

The assembly of micrometer-sized parts is an important manufacturing process; any development in it could potentially change the current manufacturing practices for micrometer-scale devices. Due to the lack of reliable microassembly techniques, these devices are often manufactured using silicon, which includes etching and depositions with little use of assembly processes. The result is the requirement of specialized manufacturing conditions with hazardous byproducts and limited applications where only simple mechanisms are allowed. Optical tweezers are non-contact type manipulators that are very suitable for assembling microparts and solve one of the most difficult problems for microassembly, which is the sticking of the physical manipulator to the micropart. Although contact type manipulators can be surface modified to be non-sticky, this involves extra preprocessing—optical tweezers do not require such additional efforts. The weakness of using optical tweezers is that the permanent assembly of parts is not possible as only very small forces can be applied. We introduce an advanced microassembly environment with the combined use of optical tweezers and a motorized microtip, where the former is used to position two parts and the latter is used to introduce deformation in the parts so that they form a strongly fitted assembly. (paper)

8. Listening to proteins and viruses with nanoaperture optical tweezers (Presentation Recording)

Gordon, Reuven

2015-08-01

This talk will present a nanoaperture tweezer approach to measure the acoustic spectra of viruses and single proteins. The approach, termed extraordinary optical Raman (EAR), shows promise for uncovering the structure and mechanical properties of nanoparticles as well as the effects of their interactions.

9. Noncontact microsurgery and micromanipulation of living cells with combined system femtosecond laser scalpel-optical tweezers

Il'ina, Inna V.; Sitnikov, Dmitry S.; Ovchinnikov, Andrey V.; Agranat, Mikhail B.; Khramova, Yulia V.; Semenova, Maria L.

2012-06-01

We report on the results of using self-developed combined laser system consisting of a femtosecond laser scalpel (Cr:Forsterite seed oscillator and a regenerative amplifier, 620 nm, 100 fs, 10 Hz) and optical tweezers (cw laser, 1064 nm) for performing noncontact laser-mediated polar body (PB) and trophectoderm (TE) biopsy of early mammalian embryos. To perform PB biopsy the femtosecond laser scalpel was initially used to drill an opening in the zona pellucida, and then the PB was extracted out of the zygote with the optical tweezers. Unlike PB biopsy, TE biopsy allows diagnosing maternally-derived as well as paternally-derived defects. Moreover, as multiple TE cells can be taken from the embryo, more reliable diagnosis can be done. TE biopsy was performed by applying laser pulses to dissect the desired amount of TE cells that had just left the zona pellucida during the hatching. Optical tweezers were then used to trap and move the dissected TE cells in a prescribed way. Laser power in optical tweezers and energy of femtosecond laser pulses were thoroughly optimized to prevent cell damage and obtain high viability rates. In conclusion, the proposed techniques of laser-based embryo biopsy enable accurate, contamination-free, simple and quick microprocessing of living cells.

10. Optical alignment and confinement of an ellipsoidal nanorod in optical tweezers: a theoretical study

Trojek, Jan; Chvátal, Lukáš; Zemánek, Pavel

2012-01-01

Roč. 29, č. 7 (2012), s. 1224-1236. ISSN 1084-7529 R&D Projects: GA ČR GA202/09/0348; GA MŠk ED0017/01/01 Institutional support: RVO:68081731 Keywords : ellipsoidal nanorod * optical tweezers * Rayleigh approximation Subject RIV: BH - Optics, Masers, Lasers Impact factor: 1.665, year: 2012

11. RBC aggregation dynamics in autologous plasma and serum studied with double-channel optical tweezers

Lee, Kisung; Danilina, Anna; Potkin, Anton; Kinnunen, Matti; Priezzhev, Alexander; Meglinski, Igor

2016-04-01

Red blood cells aggregating and disaggregating forces were measured in the autologous plasma and serum using the double-channeled optical tweezers. A significant, three-fold decrease of the both forces was observed in the serum compared to the plasma. The results of this study help to better assess the RBC aggregation mechanism.

12. Synthesis and Properties of Bis-Porphyrin Molecular Tweezers: Effects of Spacer Flexibility on Binding and Supramolecular Chirogenesis

Magnus Blom

2015-12-01

Full Text Available Ditopic binding of various dinitrogen compounds to three bisporphyrin molecular tweezers with spacers of varying conformational rigidity, incorporating the planar enediyne (1, the helical stiff stilbene (2, or the semi-rigid glycoluril motif fused to the porphyrins (3, are compared. Binding constants Ka = 104–106 M−1 reveal subtle differences between these tweezers, that are discussed in terms of porphyrin dislocation modes. Exciton coupled circular dichroism (ECCD of complexes with chiral dinitrogen guests provides experimental evidence for the conformational properties of the tweezers. The results are further supported and rationalized by conformational analysis.

13. Preparation and Photo-thermal Isomerization of Azo [2.2.1]-and Azo[2.2.3]metacyclophane Tweezers

Sawada, Tsuyoshi; Thiemann, Thies; Mataka, Shuntaro

1999-01-01

Azo[2.2.1]metacyclophane (MCP) tweezer 11 and azo[2.2.3]MCP tweezer 12 were prepared from [2.2.1]MCP 1a and [2.2.3]MCP 8, respectively, by ipso-nitration followed by reductive coupling using LiAlH_4. The photochemical and thermal isomerization of azo[2.2.n]MCP tweezers were studied by UV spectroscopy. The activation parameter (E_a) of the thermal cis-trans isomerization of 11 and 12 was determined as 81.7 kJmol^ and 90.9 kJmol^, respectively.

14. Low cost optical tweezers systems using double coil driving stepping motor to controlling sample stage

Laowattanatham, N.; Cheamanunkul, N.; Plaipichit, S.; Buranasiri, P.; Nuansri, R.

2013-06-01

In this research, the low cost optical tweezers systems using X-Y stage has been developed by using 5-phase stepping motor. By using sequential double coil driving, we can obtain the driving torque larger than using the single coil driving. The moving scale is fine resolution at 0.2 micrometer. The overall systems based on microcontroller PIC18F458 and joystick controller with LabView® graphical user interface (GUI). The mechanical damping has been included in the system for decreasing the vibrational noise. By using this method, our optical tweezers system is cheaper than the other commercial system that has been used the piezoelectric driving, and still has the same efficiency.

15. Optical Nanofiber Integrated into Optical Tweezers for In Situ Fiber Probing and Optical Binding Studies

Ivan Gusachenko

2015-07-01

Full Text Available Precise control of particle positioning is desirable in many optical propulsion and sorting applications. Here, we develop an integrated platform for particle manipulation consisting of a combined optical nanofiber and optical tweezers system. We show that consistent and reversible transmission modulations arise when individual silica microspheres are introduced to the nanofiber surface using the optical tweezers. The observed transmission changes depend on both particle and fiber diameter and can be used as a reference point for in situ nanofiber or particle size measurement. Thence, we combine scanning electron microscope (SEM size measurements with nanofiber transmission data to provide calibration for particle-based fiber assessment. This integrated optical platform provides a method for selective evanescent field manipulation of micron-sized particles and facilitates studies of optical binding and light-particle interaction dynamics.

16. Membrane tether formation from voltage-clamped outer hair cells using optical tweezers

Qian, Feng; Ermilov, Sergey A.; Murdock, David R.; Brownell, William E.; Anvari, Bahman

2004-06-01

Outer hair cells contribute an active mechanical feedback to the vibrations of the cochlear structures resulting in the high sensitivity and frequency selectivity of normal hearing. We have designed and implemented a novel experimental setup that combines optical tweezers with patch-clamp apparatus to investigate the electromechanical properties of cellular plasma membranes. A micron-size bead trapped by the optical tweezers is brought in contact with the membrane of a voltage-clamped cell, and subsequently moved away to form a plasma membrane tether. Bead displacement during tether elongation is monitored by a quadrant photodetector to obtain time-resolved measurements of the tethering force. Salient information associated with the mechanical properties of the membrane tether can thus be obtained. Tethers can be pulled from the cell membrane at different holding potentials, and the tether force response can be measured while changing transmembrane potential. Experimental results from outer hair cells and human embryonic kidney cells are presented.

17. Observation of a single-beam gradient force acoustical trap for elastic particles: acoustical tweezers

Baresch, Diego; Marchiano, Régis

2016-01-01

The ability to manipulate matter precisely is critical for the study and development of a large variety of systems. Optical tweezers are excellent tools to handle particles ranging in size from a few micrometers to hundreds of nanometers but become inefficient and damaging on larger objects. We demonstrate for the first reported time the trapping of elastic particles by the large gradient force of a single acoustical beam in three dimensions. We show that at equal power, acoustical forces overtake by 8 orders of magnitude that of optical ones on macroscopic objects. Acoustical tweezers can push, pull and accurately control both the position of the particle and the forces exerted under damage-free conditions. The large spectrum of frequencies covered by coherent ultrasonic sources will provide a wide variety of manipulation possibilities from macro- to microscopic length scales. We believe our observations improve the prospects for wider use of non-contact manipulation in biology, biophysics, microfluidics and...

18. Identification of individual biofilm-forming bacterial cells using Raman tweezers

Samek, Ota; Bernatová, Silvie; Ježek, Jan; Šiler, Martin; Šerý, Mojmir; Krzyžánek, Vladislav; Hrubanová, Kamila; Zemánek, Pavel; Holá, Veronika; Růžička, Filip

2015-05-01

A method for in vitro identification of individual bacterial cells is presented. The method is based on a combination of optical tweezers for spatial trapping of individual bacterial cells and Raman microspectroscopy for acquisition of spectral "Raman fingerprints" obtained from the trapped cell. Here, Raman spectra were taken from the biofilm-forming cells without the influence of an extracellular matrix and were compared with biofilm-negative cells. Results of principal component analyses of Raman spectra enabled us to distinguish between the two strains of Staphylococcus epidermidis. Thus, we propose that Raman tweezers can become the technique of choice for a clearer understanding of the processes involved in bacterial biofilms which constitute a highly privileged way of life for bacteria, protected from the external environment.

19. In Vivo Quantification of Peroxisome Tethering to Chloroplasts in Tobacco Epidermal Cells Using Optical Tweezers.

Gao, Hongbo; Metz, Jeremy; Teanby, Nick A; Ward, Andy D; Botchway, Stanley W; Coles, Benjamin; Pollard, Mark R; Sparkes, Imogen

2016-01-01

Peroxisomes are highly motile organelles that display a range of motions within a short time frame. In static snapshots, they can be juxtaposed to chloroplasts, which has led to the hypothesis that they are physically interacting. Here, using optical tweezers, we tested the dynamic physical interaction in vivo. Using near-infrared optical tweezers combined with TIRF microscopy, we were able to trap peroxisomes and approximate the forces involved in chloroplast association in vivo in tobacco (Nicotiana tabacum) and observed weaker tethering to additional unknown structures within the cell. We show that chloroplasts and peroxisomes are physically tethered through peroxules, a poorly described structure in plant cells. We suggest that peroxules have a novel role in maintaining peroxisome-organelle interactions in the dynamic environment. This could be important for fatty acid mobilization and photorespiration through the interaction with oil bodies and chloroplasts, highlighting a fundamentally important role for organelle interactions for essential biochemistry and physiological processes. PMID:26518344

20. Measurement of particle motion in optical tweezers embedded in a Sagnac interferometer

Galinskiy, Ivan; Salgado, Israel Rebolledo; Hautefeuille, Mathieu; Mehlig, Bernhard; Hanstorp, Dag

2015-01-01

We have constructed a counterpropagating optical tweezers setup embedded in a Sagnac interferometer in order to increase the sensitivity of position tracking for particles in the geometrical optics regime. The enhancement of the position determination using a Sagnac interferometer has previously been described theoretically by Taylor et al. [Journal of Optics 13, 044014 (2011)] for Rayleigh-regime particles trapped in an antinode of a standing wave. We have extended their theory to a case of arbitrarily-sized particles trapped with orthogonally-polarized counterpropagating beams. The working distance of the setup was sufficiently long to optically induce particle oscillations orthogonally to the axis of the tweezers with an auxiliary laser beam. Using these oscillations as a reference, we have experimentally shown that Sagnac-enhanced back focal plane interferometry is capable of providing an improvement of more than 5 times in the signal-to-background ratio, corresponding to a more than 30-fold improvement o...

1. Rapid formation of size-controllable multicellular spheroids via 3D acoustic tweezers.

Chen, Kejie; Wu, Mengxi; Guo, Feng; Li, Peng; Chan, Chung Yu; Mao, Zhangming; Li, Sixing; Ren, Liqiang; Zhang, Rui; Huang, Tony Jun

2016-07-01

The multicellular spheroid is an important 3D cell culture model for drug screening, tissue engineering, and fundamental biological research. Although several spheroid formation methods have been reported, the field still lacks high-throughput and simple fabrication methods to accelerate its adoption in drug development industry. Surface acoustic wave (SAW) based cell manipulation methods, which are known to be non-invasive, flexible, and high-throughput, have not been successfully developed for fabricating 3D cell assemblies or spheroids, due to the limited understanding on SAW-based vertical levitation. In this work, we demonstrated the capability of fabricating multicellular spheroids in the 3D acoustic tweezers platform. Our method used drag force from microstreaming to levitate cells in the vertical direction, and used radiation force from Gor'kov potential to aggregate cells in the horizontal plane. After optimizing the device geometry and input power, we demonstrated the rapid and high-throughput nature of our method by continuously fabricating more than 150 size-controllable spheroids and transferring them to Petri dishes every 30 minutes. The spheroids fabricated by our 3D acoustic tweezers can be cultured for a week with good cell viability. We further demonstrated that spheroids fabricated by this method could be used for drug testing. Unlike the 2D monolayer model, HepG2 spheroids fabricated by the 3D acoustic tweezers manifested distinct drug resistance, which matched existing reports. The 3D acoustic tweezers based method can serve as a novel bio-manufacturing tool to fabricate complex 3D cell assembles for biological research, tissue engineering, and drug development. PMID:27327102

2. Dielectrophoretic tweezers for examining particle-surface interactions within microfluidic devices

Lee, S. W.; LI, HAIBO; Bashir, Rashid

2007-01-01

The authors present dielectrophoresis (DEP)-based tweezers that can be used to characterize the interactions between a particle and the surface it is attached to, within a microfluidic device. As a proof of concept, 5.4 mu m polystyrene beads functionalized by carboxyl group were attached on a bare and poly-L-lysine functionalized oxide surface. Negative dielectrophoresis force was generated using interdigitated electrodes and the peak dielectrophoresis voltage where the beads were repelled a...

3. Measurement of particle motion in optical tweezers embedded in a Sagnac interferometer

Galinskiy, Ivan; Isaksson, Oscar; Salgado, Israel Rebolledo; Hautefeuille, Mathieu; Mehlig, Bernhard; Hanstorp, Dag

2015-01-01

We have constructed a counterpropagating optical tweezers setup embedded in a Sagnac interferometer in order to increase the sensitivity of position tracking for particles in the geometrical optics regime. Enhanced position determination using a Sagnac interferometer has previously been described theoretically by Taylor et al. [Journal of Optics 13, 044014 (2011)] for Rayleigh-regime particles trapped in an antinode of a standing wave. We have extended their theory to a case of arbitrarily-si...

4. A theoretical study of the feasibility of acoustical tweezers: Ray acoustics approach

Lee, Jungwoo; Ha, Kanglyeol; Shung, K. Kirk

2005-05-01

The optical tweezer has been found to have many biomedical applications in trapping macromolecules and cells. For the trapping mechanism, there has to be a sharp spatial change in axial optical intensity and the particle size must be much greater than the wavelength. Similar phenomenon may exist in acoustics. This work was undertaken to demonstrate theoretically that it is possible to acoustically trap particles near the focal point where most of the acoustic energy is concentrated if certain conditions are met. Acoustic force exerted on a fluid particle in ultrasonic fields is analyzed in a ray acoustics regime where the wavelength of acoustic beam is much smaller than the size of the particle. In order to apply the acoustical tweezer to manipulating macromolecules and cells whose size is in the order of a few microns or less, a prerequisite is that the ultrasound wavelength has to be much smaller than a few microns. In this paper, the analysis is therefore based on the field pattern produced by a strongly focused 100 MHz ultrasonic transducer with Gaussian intensity distribution. For the realization of acoustic trapping, negative axial radiation force has to be generated to pull a particle towards a focus. The fat particle considered for acoustic trapping in this paper has an acoustic impedance of 1.4 MRayls. The magnitude of the acoustic axial radiation force that has been calculated as the size of the fat particle is varied from 8λ to 14λ. In addition, both Fresnel coefficients at various positions are also calculated to assess the interaction of reflection and refraction and their relative contribution to the effect of the acoustical tweezer. The simulation results show that the feasibility of the acoustical tweezer depends on both the degree of acoustic impedance mismatch and the degree of focusing relative to the particle size. .

5. A journey in bioinspired supramolecular chemistry: from molecular tweezers to small molecules that target myotonic dystrophy

Zimmerman, Steven C

2016-01-01

This review summarizes part of the author’s research in the area of supramolecular chemistry, beginning with his early life influences and early career efforts in molecular recognition, especially molecular tweezers. Although designed to complex DNA, these hosts proved more applicable to the field of host–guest chemistry. This early experience and interest in intercalation ultimately led to the current efforts to develop small molecule therapeutic agents for myotonic dystrophy using a rationa...

6. Force mapping during the formation and maturation of cell adhesion sites with multiple optical tweezers

Schwingel, Melanie; Bastmeyer, Martin

2014-01-01

Focal contacts act as mechanosensors allowing cells to respond to their biomechanical environment. Force transmission through newly formed contact sites is a highly dynamic process requiring a stable link between the intracellular cytoskeleton and the extracellular environment. To simultaneously investigate cellular traction forces in several individual maturing adhesion sites within the same cell, we established a custom-built multiple trap optical tweezers setup. Beads functionalized with f...

7. Optical tweezers and surface plasmon resonance combination system based on the high numerical aperture lens

Shan, Xuchen; Zhang, Bei; Lan, Guoqiang; Wang, Yiqiao; Liu, Shugang

2015-11-01

Biology and medicine sample measurement takes an important role in the microscopic optical technology. Optical tweezer has the advantage of accurate capture and non-pollution of the sample. The SPR(surface plasmon resonance) sensor has so many advantages include high sensitivity, fast measurement, less consumption of sample and label-free detection of biological sample that the SPR sensing technique has been used for surface topography, analysis of biochemical and immune, drug screening and environmental monitoring. If they combine, they will play an important role in the biological, chemical and other subjects. The system we propose use the multi-axis cage system, by using the methods of reflection and transmiss ion to improve the space utilization. The SPR system and optical tweezer were builtup and combined in one system. The cage of multi-axis system gives full play to its accuracy, simplicity and flexibility. The size of the system is 20 * 15 * 40 cm3 and thus the sample can be replaced to switch between the optical tweezers system and the SPR system in the small space. It means that we get the refractive index of the sample and control the particle in the same system. In order to control the revolving stage, get the picture and achieve the data stored automatically, we write a LabVIEW procedure. Then according to the data from the back focal plane calculate the refractive index of the sample. By changing the slide we can trap the particle as optical tweezer, which makes us measurement and trap the sample at the same time.

8. Identification of individual biofilm-forming bacterial cells using Raman tweezers

Samek, Ota; Bernatová, Silvie; Ježek, Jan; Šiler, Martin; Šerý, Mojmír; Krzyžánek, Vladislav; Hrubanová, Kamila; Zemánek, Pavel; Holá, V.; Růžička, F.

2015-01-01

Roč. 20, č. 5 (2015), 051038:1-6. ISSN 1083-3668 R&D Projects: GA ČR GAP205/11/1687; GA MŠk(CZ) LO1212; GA MŠk ED0017/01/01 Institutional support: RVO:68081731 Keywords : Raman tweezers * Staphylococcus epidermidis * biofilm Subject RIV: BH - Optics, Masers, Lasers Impact factor: 2.859, year: 2014

9. Red blood cell membrane viscoelasticity, agglutination and zeta potential measurements with double optical tweezers

Fontes, Adriana; Fernandes, Heloise P.; Barjas-Castro, Maria L.; de Thomaz, André A.; de Ysasa Pozzo, Liliana; Barbosa, Luiz C.; Cesar, Carlos L.

2006-02-01

The red blood cell (RBC) viscoelastic membrane contains proteins and glycolproteins embedded in, or attached, to a fluid lipid bilayer and are negatively charged, which creates a repulsive electric (zeta) potential between the cells and prevents their aggregation in the blood stream. There are techniques, however, to decrease the zeta potential to allow cell agglutination which are the basis of most of the tests of antigen-antibody interactions in blood banks. This report shows the use of a double optical tweezers to measure RBC membrane viscosity, agglutination and zeta potential. In our technique one of the optical tweezers trap a silica bead that binds strongly to a RBC at the end of a RBCs rouleaux and, at the same time, acts as a pico-Newton force transducer, after calibration through its displacement from the equilibrium position. The other optical tweezers trap the RBC at the other end. To measure the membrane viscosity the optical force is measured as a function of the velocity between the RBCs. To measure the adhesion the tweezers are slowly displaced apart until the RBCs disagglutination happens. The RBC zeta potential is measured in two complimentary ways, by the force on the silica bead attached to a single RBC in response to an applied electric field, and the conventional way, by the measurement of terminal velocity of the RBC after released from the optical trap. These two measurements provide information about the RBC charges and, also, electrolytic solution properties. We believe this can improve the methods of diagnosis in blood banks.

10. Scanning a DNA molecule for bound proteins using hybrid magnetic and optical tweezers.

Marijn T J van Loenhout

Full Text Available The functional state of the genome is determined by its interactions with proteins that bind, modify, and move along the DNA. To determine the positions and binding strength of proteins localized on DNA we have developed a combined magnetic and optical tweezers apparatus that allows for both sensitive and label-free detection. A DNA loop, that acts as a scanning probe, is created by looping an optically trapped DNA tether around a DNA molecule that is held with magnetic tweezers. Upon scanning the loop along the λ-DNA molecule, EcoRI proteins were detected with ~17 nm spatial resolution. An offset of 33 ± 5 nm for the detected protein positions was found between back and forwards scans, corresponding to the size of the DNA loop and in agreement with theoretical estimates. At higher applied stretching forces, the scanning loop was able to remove bound proteins from the DNA, showing that the method is in principle also capable of measuring the binding strength of proteins to DNA with a force resolution of 0.1 pN/[Formula: see text]. The use of magnetic tweezers in this assay allows the facile preparation of many single-molecule tethers, which can be scanned one after the other, while it also allows for direct control of the supercoiling state of the DNA molecule, making it uniquely suitable to address the effects of torque on protein-DNA interactions.

11. Acoustic tweezers via sub–time-of-flight regime surface acoustic waves

Collins, David J.; Devendran, Citsabehsan; Ma, Zhichao; Ng, Jia Wei; Neild, Adrian; Ai, Ye

2016-01-01

Micrometer-scale acoustic waves are highly useful for refined optomechanical and acoustofluidic manipulation, where these fields are spatially localized along the transducer aperture but not along the acoustic propagation direction. In the case of acoustic tweezers, such a conventional acoustic standing wave results in particle and cell patterning across the entire width of a microfluidic channel, preventing selective trapping. We demonstrate the use of nanosecond-scale pulsed surface acoustic waves (SAWs) with a pulse period that is less than the time of flight between opposing transducers to generate localized time-averaged patterning regions while using conventional electrode structures. These nodal positions can be readily and arbitrarily positioned in two dimensions and within the patterning region itself through the imposition of pulse delays, frequency modulation, and phase shifts. This straightforward concept adds new spatial dimensions to which acoustic fields can be localized in SAW applications in a manner analogous to optical tweezers, including spatially selective acoustic tweezers and optical waveguides. PMID:27453940

12. Design and optimization of arrays of neodymium iron boron-based magnets for magnetic tweezers applications

We present the design methodology for arrays of neodymium iron boron (NdFeB)-based magnets for use in magnetic tweezers devices. Using finite element analysis (FEA), we optimized the geometry of the NdFeB magnet as well as the geometry of iron yokes designed to focus the magnetic fields toward the sample plane. Together, the magnets and yokes form a magnetic array which is the basis of the magnetic tweezers device. By systematically varying 15 distinct shape parameters, we determined those features that maximize the magnitude of the magnetic field gradient as well as the length scale over which the magnetic force operates. Additionally, we demonstrated that magnetic saturation of the yoke material leads to intrinsic limitations in any geometric design. Using this approach, we generated a compact and light-weight magnetic tweezers device that produces a high field gradient at the image plane in order to apply large forces to magnetic beads. We then fabricated the optimized yoke and validated the FEA by experimentally mapping the magnetic field of the device. The optimization data and iterative FEA approach outlined here will enable the streamlined design and construction of specialized instrumentation for force-sensitive microscopy

13. Design and optimization of arrays of neodymium iron boron-based magnets for magnetic tweezers applications.

Zacchia, Nicholas A; Valentine, Megan T

2015-05-01

We present the design methodology for arrays of neodymium iron boron (NdFeB)-based magnets for use in magnetic tweezers devices. Using finite element analysis (FEA), we optimized the geometry of the NdFeB magnet as well as the geometry of iron yokes designed to focus the magnetic fields toward the sample plane. Together, the magnets and yokes form a magnetic array which is the basis of the magnetic tweezers device. By systematically varying 15 distinct shape parameters, we determined those features that maximize the magnitude of the magnetic field gradient as well as the length scale over which the magnetic force operates. Additionally, we demonstrated that magnetic saturation of the yoke material leads to intrinsic limitations in any geometric design. Using this approach, we generated a compact and light-weight magnetic tweezers device that produces a high field gradient at the image plane in order to apply large forces to magnetic beads. We then fabricated the optimized yoke and validated the FEA by experimentally mapping the magnetic field of the device. The optimization data and iterative FEA approach outlined here will enable the streamlined design and construction of specialized instrumentation for force-sensitive microscopy. PMID:26026529

14. Optical macro-tweezers: trapping of highly motile micro-organisms

Optical micromanipulation stands for contact-free handling of microscopic particles by light. Optical forces can manipulate non-absorbing objects in a large range of sizes, e.g., from biological cells down to cold atoms. Recently much progress has been made going from the micro- down to the nanoscale. Less attention has been paid to going the other way, trapping increasingly large particles. Optical tweezers typically employ a single laser beam tightly focused by a microscope objective of high numerical aperture to stably trap a particle in three dimensions (3D). As the particle size increases, stable 3D trapping in a single-beam trap requires scaling up the optical power, which eventually induces adverse biological effects. Moreover, the restricted field of view of standard optical tweezers, dictated by the use of high NA objectives, is particularly unfavorable for catching actively moving specimens. Both problems can be overcome by traps with counter-propagating beams. Our 'macro-tweezers' are especially designed to trap highly motile organisms, as they enable three-dimensional all-optical trapping and guiding in a volume of 2 × 1 × 2 mm3. Here we report for the first time the optical trapping of large actively swimming organisms, such as for instance Euglena protists and dinoflagellates of up to 70 µm length. Adverse bio-effects are kept low since trapping occurs outside high intensity regions, e.g., focal spots. We expect our approach to open various possibilities in the contact-free handling of 50–100 µm sized objects that could hitherto not be envisaged, for instance all-optical holding of individual micro-organisms for taxonomic identification, selective collecting or tagging

15. tweezercalib 2.0: Faster version of MatLab package for precise calibration of optical tweezers

Hansen, Poul Martin; Tolic-Nørrelykke, Iva Marija; Flyvbjerg, Henrik;

2006-01-01

of factors that affect this power spectrum, as described in vs. 1 of the package. The graphical user interface allows the user to include of leave out each of these factors. Several "health tests" are applied to the experimental data during calibration and test results are displayed graphically. Thus......We present a vectorized version of the MatLab (MathWorks Inc) package tweezercalib for calibration of optical tweezers with precision. The calibration is based on the power spectrum of the Brownian motion of a dielectric bead trapped in the tweezers. Precision is achieved by accounting for a number...

16. Measurement of Breaking Force of Fluorescence Labelled Microtubules with Optical Tweezers

LIU Chun-Xiang; GUO Hong-Lian; XU Chun-Hua; YUAN Ming; LI Znao-Lin; CHENG Bing-Ying; ZHANG Dao-Zhong

2005-01-01

@@ Under illumination of excitation light, the force that can make fluorescent dye-labelled microtubules break up is measured by using dual-beam optical tweezers. It is found that this force is about several piconewtons, which is two orders of magnitude smaller than that without fluorescence label. Microtubules can be elongated about 20% and the increase of the tensile force is nonlinear with the microtubule elongation. Some qualitative explanations are given for the mechanisms about the breakup and elongation of microtubules exposed to excitation light.

17. A journey in bioinspired supramolecular chemistry: from molecular tweezers to small molecules that target myotonic dystrophy

2016-01-01

Summary This review summarizes part of the author’s research in the area of supramolecular chemistry, beginning with his early life influences and early career efforts in molecular recognition, especially molecular tweezers. Although designed to complex DNA, these hosts proved more applicable to the field of host–guest chemistry. This early experience and interest in intercalation ultimately led to the current efforts to develop small molecule therapeutic agents for myotonic dystrophy using a rational design approach that heavily relies on principles of supramolecular chemistry. How this work was influenced by that of others in the field and the evolution of each area of research is highlighted with selected examples. PMID:26877815

18. Template stripped double nanohole in a gold film for nano-optical tweezers

Double nanohole (DNH) laser tweezers can optically trap and manipulate objects such as proteins, nanospheres, and other nanoparticles; however, precise fabrication of those DNHs has been expensive with low throughput. In this work, template stripping was used to pattern DNHs with gaps as small as 7 nm, in optically thick Au films. These DNHs were used to trap streptavidin as proof of operation. The structures were processed multiple times from the same template to demonstrate reusability. Template stripping is a promising method for high-throughput, reproducible, and cost efficient fabrication of DNH apertures for optical trapping. (paper)

19. Fast acoustic tweezers for the two-dimensional manipulation of individual particles in microfluidic channels

Tran, S B Q; Thibault, Pierre; 10.1063/1.4751348

2012-01-01

This paper presents a microfluidic device that implements standing surface acoustic waves in order to handle single cells, droplets, and generally particles. The particles are moved in a very controlled manner by the two-dimensional drifting of a standing wave array, using a slight frequency modulation of two ultrasound emitters around their resonance. These acoustic tweezers allow any type of motion at velocities up to few 10mm/s, while the device transparency is adapted for optical studies. The possibility of automation provides a critical step in the development of lab-on-a-chip cell sorters and it should find applications in biology, chemistry, and engineering domains.

20. Quantitative characterization for dielectrophoretic behavior of biological cells using optical tweezers

Park, In Soo; Hee Park, Se; Woo Lee, Sang; Sung Yoon, Dae; Kim, Beop-Min

2014-02-01

We report a method to precisely quantify dielectrophoretic (DEP) forces and cutoff frequencies (fc) of viable and nonviable yeast cells. The method consists of a two-step process in which generated DEP forces act upon a cell through a micro-electrode device, followed by direct measurement of DEP forces using optical tweezers. DEP behaviors of viable and nonviable yeast cells are monitored as a function of AC frequency. We believe that the proposed method can be used as a powerful platform for cell-based assays to characterize the DEP behavior of various cell types including cancer and normal cells.

1. Laser-induced fusion of human embryonic stem cells with optical tweezers

Chen Shuxun; Wang Xiaolin; Sun Dong [Department of Mechanical and Biomedical Engineering, City University of Hong Kong (Hong Kong); Cheng Jinping; Han Cheng, Shuk [Department of Biology and Chemistry, City University of Hong Kong (Hong Kong); Kong, Chi-Wing [Stem Cell and Regenerative Medicine Consortium, and Departments of Medicine and Physiology, LKS Faculty of Medicine, University of Hong Kong (Hong Kong); Li, Ronald A. [Stem Cell and Regenerative Medicine Consortium, and Departments of Medicine and Physiology, LKS Faculty of Medicine, University of Hong Kong (Hong Kong); Center of Cardiovascular Research, Mount Sinai School of Medicine, New York, New York 10029 (United States)

2013-07-15

We report a study on the laser-induced fusion of human embryonic stem cells (hESCs) at the single-cell level. Cells were manipulated by optical tweezers and fused under irradiation with pulsed UV laser at 355 nm. Successful fusion was indicated by green fluorescence protein transfer. The influence of laser pulse energy on the fusion efficiency was investigated. The fused products were viable as gauged by live cell staining. Successful fusion of hESCs with somatic cells was also demonstrated. The reported fusion outcome may facilitate studies of cell differentiation, maturation, and reprogramming.

2. Data on force-dependent structural changes of chromatin fibers measured with magnetic tweezers

Fan-Tso Chien

2014-12-01

Full Text Available The compaction of chromatin fibers regulates the accessibility of embedded DNA, highly associated with transcriptional activities [1]. Single molecule force spectroscopy has revealed the great details of the structural changes of chromatin fibers in the presence of external exerted force [2–7]. However, most of the studies focus on a specific force regime [2,3,8,9]. The data here show force-extension (FE traces of chromatin fibers as measured with magnetic tweezers, covering the force regime from 0 pN to 27 pN. Those traces provide information for further studies at varied force regimes.

3. Optical tweezers study of red blood cell aggregation and disaggregation in plasma and protein solutions

Lee, Kisung; Kinnunen, Matti; Khokhlova, Maria D.; Lyubin, Evgeny V.; Priezzhev, Alexander V.; Meglinski, Igor; Fedyanin, Andrey A.

2016-03-01

Kinetics of optical tweezers (OT)-induced spontaneous aggregation and disaggregation of red blood cells (RBCs) were studied at the level of cell doublets to assess RBC interaction mechanics. Measurements were performed under in vitro conditions in plasma and fibrinogen and fibrinogen + albumin solutions. The RBC spontaneous aggregation kinetics was found to exhibit different behavior depending on the cell environment. In contrast, the RBC disaggregation kinetics was similar in all solutions qualitatively and quantitatively, demonstrating a significant contribution of the studied proteins to the process. The impact of the study on assessing RBC interaction mechanics and the protein contribution to the reversible RBC aggregation process is discussed.

4. Formation of an artificial blood vessel: adhesion force measurements with optical tweezers

Knoener, Gregor; Campbell, Julie H.; Heckenberg, Norman R.; Rubinsztein-Dunlop, Halina

2004-10-01

We are investigating the formation of a tissue capsule around a foreign body. This tissue capsule can be used as an autologous graft for the replacement of diseased blood vessels or for bypass surgery. The graft is grown in the peritoneal cavity of the recipient and the formation starts with the adhesion of cells to the foreign body. We identify the cell type and measure the adhesion of these cells to foreign materials using optical tweezers. Cell adhesion to macroscopic samples and microspheres is investigated. No difference in the adhesion force was measurable for polyethylene, silicon and Tygon on a scale accessible to optical tweezers. The density of adherent cells was found to vary strongly, being highest on polyethylene. The mean rupture forces for cell-microsphere adhesion ranged from 24 to 39 pN and changed upon preadsorption of bovine serum albumin. For plain microspheres, the highest mean rupture force was found for PMMA, which also showed the highest adhesion probability for the cell-microsphere interaction.

5. Optical tweezers and non-ratiometric fluorescent-dye-based studies of respiration in sperm mitochondria

The purpose of this study is to investigate how the mitochondrial membrane potential affects sperm motility using laser tweezers and a non-ratiometric fluorescent probe, DiOC6(3). A 1064 nm Nd:YVO4 continuous wave laser was used to trap motile sperm at a power of 450 mW in the trap spot. Using customized tracking software, the curvilinear velocity (VCL) and the escape force from the laser tweezers were measured. Human (Homo sapiens), dog (Canis lupis familiaris) and drill (Mandrillus leucophaeus) sperm were treated with DiOC6(3) to measure the membrane potential in the mitochondria-rich sperm midpieces. Sperm from all three species exhibited an increase in fluorescence when treated with the DiOC6(3). When a cyanide inhibitor (CCCP) of aerobic respiration was applied, sperm of all three species exhibited a reduction in fluorescence to pre-dye levels. With respect to VCL and escape force, the CCCP had no effect on dog or human sperm, suggesting a major reliance upon anaerobic respiration (glycolysis) for ATP in these two species. Based on the preliminary study on drill sperm, CCCP caused a drop in the VCL, suggesting potential reliance on both glycolysis and aerobic respiration for motility. The results demonstrate that optical trapping in combination with DiOC6(3) is an effective way to study sperm motility and energetics

6. Counter-propagating dual-trap optical tweezers based on linear momentum conservation

Ribezzi-Crivellari, M.; Huguet, J. M. [Small Biosystems Lab, Dept. de Fisica Fonamental, Universitat de Barcelona, Avda. Diagonal 647, 08028 Barcelona (Spain); Ritort, F. [Small Biosystems Lab, Dept. de Fisica Fonamental, Universitat de Barcelona, Avda. Diagonal 647, 08028 Barcelona (Spain); Ciber-BBN de Bioingenieria, Biomateriales y Nanomedicina, Instituto de Salud Carlos III, Madrid (Spain)

2013-04-15

We present a dual-trap optical tweezers setup which directly measures forces using linear momentum conservation. The setup uses a counter-propagating geometry, which allows momentum measurement on each beam separately. The experimental advantages of this setup include low drift due to all-optical manipulation, and a robust calibration (independent of the features of the trapped object or buffer medium) due to the force measurement method. Although this design does not attain the high-resolution of some co-propagating setups, we show that it can be used to perform different single molecule measurements: fluctuation-based molecular stiffness characterization at different forces and hopping experiments on molecular hairpins. Remarkably, in our setup it is possible to manipulate very short tethers (such as molecular hairpins with short handles) down to the limit where beads are almost in contact. The setup is used to illustrate a novel method for measuring the stiffness of optical traps and tethers on the basis of equilibrium force fluctuations, i.e., without the need of measuring the force vs molecular extension curve. This method is of general interest for dual trap optical tweezers setups and can be extended to setups which do not directly measure forces.

7. A theoretical study of the feasibility of acoustical tweezer: Ray acoustics approach

Lee, Jungwoo; Shung, Kirk

2005-04-01

Optical tweezer has been found to have many biomedical applications in trapping macromolecules and cells. For the trapping mechanism, there has to be a sharp spatial change in axial optical intensity and the particle size must be much greater than the wavelength. Similar phenomenon may exist in acoustics. This work was undertaken to demonstrate theoretically that it is possible to acoustically trap particles near the focal point if certain conditions are met. Acoustic force exerted on fat tissue in ultrasonic fields is analyzed in ray acoustics regime where the wavelength of acoustic beam is much smaller than the size of the particle. In this paper, the analysis is therefore based on the field pattern produced by a strongly focused 100 MHz ultrasonic transducer with Gaussian intensity distribution. The magnitude of force and Fresnel coefficients at various positions are calculated. According to the simulation results, acoustical tweezer works particularly when the beam width at focus is one wavelength and the tolerance of acoustic impedance mismatch between two media lies within 6.7%. [Work supported by NIH Grant P41-EB2182.

8. High-resolution dual-trap optical tweezers with differential detection: alignment of instrument components.

Bustamante, Carlos; Chemla, Yann R; Moffitt, Jeffrey R

2009-10-01

Optical traps or "optical tweezers" have become an indispensable tool in understanding fundamental biological processes. Using our design, a dual-trap optical tweezers with differential detection, we can detect length changes to a DNA molecule tethering the trapped beads of 1 bp. By forming two traps from the same laser and maximizing the common optical paths of the two trapping beams, we decouple the instrument from many sources of environmental and instrumental noise that typically limit spatial resolution. The performance of a high-resolution instrument--the formation of strong traps, the minimization of background signals from trap movements, or the mitigation of the axial coupling, for example--can be greatly improved through careful alignment. This procedure, which is described in this article, starts from the laser and advances through the instrument, component by component. Alignment is complicated by the fact that the trapping light is in the near infrared (NIR) spectrum. Standard infrared viewing cards are commonly used to locate the beam, but unfortunately, bleach quickly. As an alternative, we use an IR-viewing charge-coupled device (CCD) camera equipped with a C-mount telephoto lens and display its image on a monitor. By visualizing the scattered light on a pair of irises of identical height separated by >12 in., the beam direction can be set very accurately along a fixed axis. PMID:20147041

9. A study of red blood cell deformability in diabetic retinopathy using optical tweezers

Smart, Thomas J.; Richards, Christopher J.; Bhatnagar, Rhythm; Pavesio, Carlos; Agrawal, Rupesh; Jones, Philip H.

2015-08-01

Diabetic retinopathy (DR) is a microvascular complication of diabetes mellitus (DM) in which high blood sugar levels cause swelling, leaking and occlusions in the blood vessels of the retina, often resulting in a loss of sight. The microvascular system requires red blood cells (RBCs) to undergo significant cellular deformation in order to pass through vessels whose diameters are significantly smaller than their own. There is evidence to suggest that DM impairs the deformability of RBCs, and this loss of deformability has been associated with diabetic kidney disease (or nephropathy) - another microvascular complication of DM. However, it remains unclear whether reduced deformability of RBCs correlates with the presence of DR. Here we present an investigation into the deformability of RBCs in patients with diabetic retinopathy using optical tweezers. To extract a value for the deformability of RBCs we use a dual-trap optical tweezers set-up to stretch individual RBCs. RBCs are trapped directly (i.e. without micro-bead handles), so rotate to assume a `side-on' orientation. Video microscopy is used to record the deformation events, and shape analysis software is used to determine parameters such as initial and maximum RBC length, allowing us to calculate the deformability for each RBC. A small decrease in deformability of diabetes cells subject to this stretching protocol is observed when compared to control cells. We also report on initial results on three dimensional imaging of individual RBCs using defocussing microscopy.

10. Single-fiber tweezers applied for dye lasing in a fluid droplet.

Liu, Zhihai; Chen, Yunhao; Zhao, Li; Zhang, Yu; Wei, Yong; Li, Hanyang; Liu, Yongjun; Zhang, Yaxun; Zhao, Enming; Yang, Xinghua; Zhang, Jianzhong; Yuan, Libo

2016-07-01

We report on the first demonstration of a single-fiber optical tweezer that is utilized to stabilize and control the liquid droplet for dye lasing. In order to trap a liquid droplet with a diameter of 15-30 μm, an annular core micro-structured optical fiber is adopted. By using wavelength division multiplexing technology, we couple a trapping light source (980 nm) and a pumping light source (532 nm) into the annular core of the fiber to realize the trapping, controlling, and pumping of the oil droplet. We show that the laser emission spectrum tunes along the same size as the oil droplet. The lasing threshold of the oil droplet with the diameter of 24 μm is 0.7 μJ. The presented fiber-based optical manipulation of liquid droplet micro-lasers can be easily combined with the micro-fluidic chip technology and also may extend the application of optical fiber tweezers for micro-droplet lasing technology in the biological field. PMID:27367077

11. Probing Mechanical Properties of Jurkat Cells under the Effect of ART Using Oscillating Optical Tweezers

2015-01-01

Acute lymphoid leukemia is a common type of blood cancer and chemotherapy is the initial treatment of choice. Quantifying the effect of a chemotherapeutic drug at the cellular level plays an important role in the process of the treatment. In this study, an oscillating optical tweezer was employed to characterize the frequency-dependent mechanical properties of Jurkat cells exposed to the chemotherapeutic agent, artesunate (ART). A motion equation for a bead bound to a cell was applied to describe the mechanical characteristics of the cell cytoskeleton. By comparing between the modeling results and experimental results from the optical tweezer, the stiffness and viscosity of the Jurkat cells before and after the ART treatment were obtained. The results demonstrate a weak power-law dependency of cell stiffness with frequency. Furthermore, the stiffness and viscosity were increased after the treatment. Therefore, the cytoskeleton cell stiffness as the well as power-law coefficient can provide a useful insight into the chemo-mechanical relationship of drug treated cancer cells and may serve as another tool for evaluating therapeutic performance quantitatively. PMID:25928073

12. Measurement of particle motion in optical tweezers embedded in a Sagnac interferometer.

Galinskiy, Ivan; Isaksson, Oscar; Salgado, Israel Rebolledo; Hautefeuille, Mathieu; Mehlig, Bernhard; Hanstorp, Dag

2015-10-19

We have constructed a counterpropagating optical tweezers setup embedded in a Sagnac interferometer in order to increase the sensitivity of position tracking for particles in the geometrical optics regime. Enhanced position determination using a Sagnac interferometer has previously been described theoretically by Taylor et al. [Journal of Optics 13, 044014 (2011)] for Rayleigh-regime particles trapped in an antinode of a standing wave. We have extended their theory to a case of arbitrarily-sized particles trapped with orthogonally-polarized counter-propagating beams. The working distance of the setup was sufficiently long to optically induce particle oscillations orthogonally to the axis of the tweezers with an auxiliary laser beam. Using these oscillations as a reference, we have experimentally shown that Sagnac-enhanced back focal plane interferometry is capable of providing an improvement of more than 5 times in the signal-to-background ratio, corresponding to a more than 30-fold improvement of the signal-to-noise ratio. The experimental results obtained are consistent with our theoretical predictions. In the experimental setup, we used a method of optical levitator-assisted liquid droplet delivery in air based on commercial inkjet technology, with a novel method to precisely control the size of droplets. PMID:26480368

13. Crosstalk elimination in the detection of dual-beam optical tweezers by spatial filtering

Ott, Dino; Oddershede, Lene B., E-mail: oddershede@nbi.dk [Niels Bohr Institute (NBI), University of Copenhagen, Blegdamsvej 17, DK-2100 Copenhagen (Denmark); Reihani, S. Nader S. [Department of Physics, Sharif University of Technology, 11369-9161 Tehran (Iran, Islamic Republic of)

2014-05-15

In dual-beam optical tweezers, the accuracy of position and force measurements is often compromised by crosstalk between the two detected signals, this crosstalk leading to systematic and significant errors on the measured forces and distances. This is true both for dual-beam optical traps where the splitting of the two traps is done by polarization optics and for dual optical traps constructed by other methods, e.g., holographic tweezers. If the two traps are orthogonally polarized, most often crosstalk is minimized by inserting polarization optics in front of the detector; however, this method is not perfect because of the de-polarization of the trapping beam introduced by the required high numerical aperture optics. Here we present a simple and easy-to-implement method to efficiently eliminate crosstalk. The method is based on spatial filtering by simply inserting a pinhole at the correct position and is highly compatible with standard back focal plane photodiode based detection of position and force. Our spatial filtering method reduces crosstalk up to five times better than polarization filtering alone. The effectiveness is dependent on pinhole size and distance between the traps and is here quantified experimentally and reproduced by theoretical modeling. The method here proposed will improve the accuracy of force-distance measurements, e.g., of single molecules, performed by dual-beam optical traps and hence give much more scientific value for the experimental efforts.

14. Crosstalk elimination in the detection of dual-beam optical tweezers by spatial filtering

Ott, Dino; Reihani, S. Nader S.; Oddershede, Lene B.

2014-05-01

In dual-beam optical tweezers, the accuracy of position and force measurements is often compromised by crosstalk between the two detected signals, this crosstalk leading to systematic and significant errors on the measured forces and distances. This is true both for dual-beam optical traps where the splitting of the two traps is done by polarization optics and for dual optical traps constructed by other methods, e.g., holographic tweezers. If the two traps are orthogonally polarized, most often crosstalk is minimized by inserting polarization optics in front of the detector; however, this method is not perfect because of the de-polarization of the trapping beam introduced by the required high numerical aperture optics. Here we present a simple and easy-to-implement method to efficiently eliminate crosstalk. The method is based on spatial filtering by simply inserting a pinhole at the correct position and is highly compatible with standard back focal plane photodiode based detection of position and force. Our spatial filtering method reduces crosstalk up to five times better than polarization filtering alone. The effectiveness is dependent on pinhole size and distance between the traps and is here quantified experimentally and reproduced by theoretical modeling. The method here proposed will improve the accuracy of force-distance measurements, e.g., of single molecules, performed by dual-beam optical traps and hence give much more scientific value for the experimental efforts.

15. Enhanced cell sorting and manipulation with combined optical tweezer and microfluidic chip technologies.

Wang, Xiaolin; Chen, Shuxun; Kong, Marco; Wang, Zuankai; Costa, Kevin D; Li, Ronald A; Sun, Dong

2011-11-01

Sorting (or isolation) and manipulation of rare cells with high recovery rate and purity are of critical importance to a wide range of physiological applications. In the current paper, we report on a generic single cell manipulation tool that integrates optical tweezers and microfluidic chip technologies for handling small cell population sorting with high accuracy. The laminar flow nature of microfluidics enables the targeted cells to be focused on a desired area for cell isolation. To recognize the target cells, we develop an image processing methodology with a recognition capability of multiple features, e.g., cell size and fluorescence label. The target cells can be moved precisely by optical tweezers to the desired destination in a noninvasive manner. The unique advantages of this sorter are its high recovery rate and purity in small cell population sorting. The design is based on dynamic fluid and dynamic light pattern, in which single as well as multiple laser traps are employed for cell transportation, and a recognition capability of multiple cell features. Experiments of sorting yeast cells and human embryonic stem cells are performed to demonstrate the effectiveness of the proposed cell sorting approach. PMID:21918752

16. Probing mechanical properties of Jurkat cells under the effect of ART using oscillating optical tweezers.

Samaneh Khakshour

Full Text Available Acute lymphoid leukemia is a common type of blood cancer and chemotherapy is the initial treatment of choice. Quantifying the effect of a chemotherapeutic drug at the cellular level plays an important role in the process of the treatment. In this study, an oscillating optical tweezer was employed to characterize the frequency-dependent mechanical properties of Jurkat cells exposed to the chemotherapeutic agent, artesunate (ART. A motion equation for a bead bound to a cell was applied to describe the mechanical characteristics of the cell cytoskeleton. By comparing between the modeling results and experimental results from the optical tweezer, the stiffness and viscosity of the Jurkat cells before and after the ART treatment were obtained. The results demonstrate a weak power-law dependency of cell stiffness with frequency. Furthermore, the stiffness and viscosity were increased after the treatment. Therefore, the cytoskeleton cell stiffness as the well as power-law coefficient can provide a useful insight into the chemo-mechanical relationship of drug treated cancer cells and may serve as another tool for evaluating therapeutic performance quantitatively.

17. Holographic optical tweezers: microassembling of shape-complementary 2PP building blocks

Ksouri, Sarah Isabelle; Mattern, Manuel; Köhler, Jannis; Aumann, Andreas; Zyla, Gordon; Ostendorf, Andreas

2014-09-01

Based on an ongoing trend in miniaturization and due to the increased complexity in MEMS-technology new methods of assembly need to be developed. Recent developments show that particularly optical forces are suitable to meet the requirements. The unique advantages of optical tweezers (OT) are attractive due to their contactless and precise manipulation forces. Spherical as well as non-spherical shaped pre-forms can already be assembled arbitrarily by using appropriate beam profiles generated by a spatial light modulator (SLM), resulting in a so called holographic optical tweezer (HOT) setup. For the fabrication of shape-complementary pre-forms, a two-photon-polymerization (2PP) process is implemented. The purpose of the process combination of 2PP and HOT is the development of an optical microprocessing platform for assembling arbitrary building blocks. Here, the optimization of the 2PP and HOT processes is described in order to allow the fabrication and 3D assembling of interlocking components. Results include the analysis of the dependence of low and high qualities of 2PP microstructures and their manufacturing accuracy for further HOT assembling processes. Besides, the applied detachable interlocking connections of the 2PP building blocks are visualized by an application example. In the long-term a full optical assembly method without applying any mechanical forces can thus be realized.

18. Chemotaxis study using optical tweezers to observe the strength and directionality of forces of Leishmania amazonensis

Pozzo, Liliana d. Y.; Fontes, Adriana; de Thomaz, André A.; Barbosa, Luiz C.; Ayres, Diana C.; Giorgio, Selma; Cesar, Carlos L.

2006-08-01

The displacements of a dielectric microspheres trapped by an optical tweezers (OT) can be used as a force transducer for mechanical measurements in life sciences. This system can measure forces on the 50 femto Newtons to 200 pico Newtons range, of the same order of magnitude of a typical forces induced by flagellar motion. The process in which living microorganisms search for food and run away from poison chemicals is known is chemotaxy. Optical tweezers can be used to obtain a better understanding of chemotaxy by observing the force response of the microorganism when placed in a gradient of attractors and or repelling chemicals. This report shows such observations for the protozoa Leishmania amazomenzis, responsible for the leishmaniasis, a serious tropical disease. We used a quadrant detector to monitor the movement of the protozoa for different chemicals gradient. This way we have been able to observe both the force strength and its directionality. The characterization of the chemotaxis of these parasites can help to understand the infection mechanics and improve the diagnosis and the treatments employed for this disease.

19. Application of optical tweezers and excimer laser to study protoplast fusion

Kantawang, Titirat; Samipak, Sompid; Limtrakul, Jumras; Chattham, Nattaporn

2015-07-01

Protoplast fusion is a physical phenomenon that two protoplasts come in contact and fuse together. Doing so, it is possible to combine specific genes from one protoplast to another during fusion such as drought resistance and disease resistance. There are a few possible methods to induce protoplast fusion, for example, electrofusion and chemical fusion. In this study, chemical fusion was performed with laser applied as an external force to enhance rate of fusion and observed under a microscope. Optical tweezers (1064 nm with 100X objective N.A. 1.3) and excimer laser (308 nm LMU-40X-UVB objective) were set with a Nikon Ti-U inverted microscope. Samples were prepared by soaking in hypertonic solution in order to induce cell plasmolysis. Elodea Canadensis and Allium cepa plasmolysed leaves were cut and observed under microscope. Concentration of solution was varied to induce difference turgor pressures on protoplasts pushing at cell wall. Free protoplasts in solution were trapped by optical tweezers to study the effect of Polyethylene glycol (PEG) solution. PEG was diluted by Ca+ solution during the process to induced protoplast cell contact and fusion. Possibility of protoplast fusion by excimer laser was investigated and found possible. Here we report a novel tool for plant cell fusion using excimer laser. Plant growth after cell fusion is currently conducted.

20. Luminescent nanoparticle trapping with far-field optical fiber-tip tweezers.

Decombe, Jean-Baptiste; Valdivia-Valero, Francisco J; Dantelle, Géraldine; Leménager, Godefroy; Gacoin, Thierry; Colas des Francs, Gérard; Huant, Serge; Fick, Jochen

2016-03-01

We report stable and reproducible trapping of luminescent dielectric YAG:Ce(3+) nanoparticles with sizes down to 60 nm using far-field dual fiber tip optical tweezers. The particles are synthesized by a specific glycothermal route followed by an original protected annealing step, resulting in significantly enhanced photostability. The tweezers properties are analyzed by studying the trapped particles residual Brownian motion using video or reflected signal records. The trapping potential is harmonic in the transverse direction to the fiber axis, but reveals interference fringes in the axial direction. Large trapping stiffness of 35 and 2 pN μm(-1) W(-1) is measured for a fiber tip-to-tip distance of 3 μm and 300 nm and 60 nm particles, respectively. The forces acting on the nanoparticles are discussed within the dipolar approximation (gradient and scattering force contributions) or exact calculations using the Maxwell Stress Tensor formalism. Prospects for trapping even smaller particles are discussed. PMID:26883602

1. The integration of a micropipette in a closed microfluidic chip with optical tweezers for investigations of single cells: erratum

Alrifaiy, Ahmed; Ramser, Kerstin

2012-01-01

In July 2011 a new concept of a closed microfluidic system equipped with a fixed micropipette, optical tweezers and a UV-Vis spectrometer was presented [Biomed. Opt. Express 2, 2299 (2011)]. Figure 1 showed falsely oriented mirrors. To clarify the design of the setup, this erratum presents a correct schematic.

2. An optical tweezer-based study of antimicrobial activity of silver nanoparticles

Yogesha; Sarbari Bhattacharya; M K Rabinal; Sharath Ananthamurthy

2012-08-01

Understanding and characterizing microbial activity reduction in the presence of antimicrobial agents can help in the design and manufacture of antimicrobial drugs. We demonstrate the use of an optical tweezer setup in recording the changes in bacterial activity with time, induced by the presence of foreign bodies in a bacterial suspension. This is achieved by monitoring the fluctuations of an optically trapped polystyrene bead immersed in it. Examining the changes in the fluctuation pattern of the bead with time provides an accurate characterization of the reduction in the microbial activity. Here, we report on the effect of addition of silver nanoparticles on bacterial cultures of Pseudomonas aeroginosa, Escherichia coli and Bacillus subtilis. We observe a decrease in the bacterial activity with time for the investigated bacterial samples. This method in our opinion, enables one to track changes in bacterial activity levels as a function of time of contact with the antibacterial agent with greater efficacy than traditional cell counting methods.

3. Rapid feedback control and stabilization of an optical tweezers with a budget microcontroller

Nino, Daniel; Wang, Haowei; Milstein, Joshua N.

2014-09-01

Laboratories ranging the scientific disciplines employ feedback control to regulate variables within their experiments, from the flow of liquids within a microfluidic device to the temperature within a cell incubator. We have built an inexpensive, yet fast and rapidly deployed, feedback control system that is straightforward and flexible to implement from a commercially available Arduino Due microcontroller. This is in comparison with the complex, time-consuming and often expensive electronics that are commonly implemented. As an example of its utility, we apply our feedback controller to the task of stabilizing the main trapping laser of an optical tweezers. The feedback controller, which is inexpensive yet fast and rapidly deployed, was implemented from hacking an open source Arduino Due microcontroller. Our microcontroller based feedback system can stabilize the laser intensity to a few tenths of a per cent at 200 kHz, which is an order of magnitude better than the laser's base specifications, illustrating the utility of these devices.

4. Robert Feulgen Prize Lecture. Laser tweezers and multiphoton microscopes in life sciences.

König, K

2000-08-01

Near infrared (NIR) laser microscopy enables optical micromanipulation, piconewton force determination, and sensitive fluorescence studies by laser tweezers. Otherwise, fluorescence images with high spatial and temporal resolution of living cells and tissues can be obtained via non-resonant fluorophore excitation with multiphoton NIR laser scanning microscopes. Furthermore, NIR femtosecond laser pulses at TW/cm2 intensities can be used to realize non-invasive contact-free surgery of nanometer-sized structures within living cells and tissues. Applications of these novel versatile NIR laser-based tools for the determination of motility forces, coenzyme and chlorophyll imaging, three-dimensional multigene detection, non-invasive optical sectioning of tissues ("optical biopsy"), functional protein imaging, and nanosurgery of chromosomes are described. PMID:11052257

5. Nanoplasmonic Tweezers Visualize Protein p53 Suppressing Unzipping of Single DNA-Hairpins

Kotnala, Abhay

2014-01-01

Here we report on the use of double-nanohole (DNH) optical tweezers as a label-free and free-solution single-molecule probe for protein-DNA interactions. Using this approach, we demonstrate the unzipping of individual 10 base pair DNA-hairpins, and quantify how tumor suppressor p53 protein delays the unzipping. From the Arrhenius behavior, we find the energy barrier to unzipping introduced by p53 to be $2\\times 10^{-20}$ J, whereas cys135ser mutant p53 does not show suppression of unzipping, which gives clues to its functional inability to suppress tumor growth. This transformative approach to single molecule analysis allows for ultra-sensitive detection and quantification of protein-DNA interactions to revolutionize the fight against genetic diseases.

6. Micro-particle manipulation by single beam acoustic tweezers based on hydrothermal PZT thick film

Benpeng Zhu

2016-03-01

Full Text Available Single-beam acoustic tweezers (SBAT, used in laboratory-on-a-chip (LOC device has promising implications for an individual micro-particle contactless manipulation. In this study, a freestanding hydrothermal PZT thick film with excellent piezoelectric property (d33 = 270pC/N and kt = 0.51 was employed for SBAT applications and a press-focusing technology was introduced. The obtained SBAT, acting at an operational frequency of 50MHz, a low f-number (∼0.9, demonstrated the capability to trap and manipulate a micro-particle sized 10μm in the distilled water. These results suggest that such a device has great potential as a manipulator for a wide range of biomedical and chemical science applications.

7. Optically-driven red blood cell rotor in linearly polarized laser tweezers

Manas Khan; Samarendra K Mohanty; A K Sood

2005-11-01

We have constructed a dual trap optical tweezers set-up around an inverted microscope where both the traps can be independently controlled and manipulated in all the three dimensions. Here we report our observations on rotation of red blood cells (RBCs) in a linearly polarized optical trap. Red blood cells deform and become twisted in hypertonic phosphate buffer saline and when trapped, experience an unbalanced radiation pressure force. The torque generated from the unbalanced force causes the trapped RBC to rotate. Addition of Ca++ ions in the solution, keeping the osmolarity same, makes the cell membranes stiffer and the cells deform less. Thus the speed of rotation of the red blood cells can be controlled, as less deformation and in turn less asymmetry in shape produces less torque under the radiation pressure resulting in slower rotation at the same laser power.

8. Microfluidic platform combining droplets and magnetic tweezers: application to HER2 expression in cancer diagnosis.

Ferraro, Davide; Champ, Jérôme; Teste, Bruno; Serra, Marco; Malaquin, Laurent; Viovy, Jean-Louis; de Cremoux, Patricia; Descroix, Stephanie

2016-01-01

The development of precision medicine, together with the multiplication of targeted therapies and associated molecular biomarkers, call for major progress in genetic analysis methods, allowing increased multiplexing and the implementation of more complex decision trees, without cost increase or loss of robustness. We present a platform combining droplet microfluidics and magnetic tweezers, performing RNA purification, reverse transcription and amplification in a fully automated and programmable way, in droplets of 250nL directly sampled from a microtiter-plate. This platform decreases sample consumption about 100 fold as compared to current robotized platforms and it reduces human manipulations and contamination risk. The platform's performance was first evaluated on cell lines, showing robust operation on RNA quantities corresponding to less than one cell, and then clinically validated with a cohort of 21 breast cancer samples, for the determination of their HER2 expression status, in a blind comparison with an established routine clinical analysis. PMID:27157697

9. Rapid feedback control and stabilization of an optical tweezers with a budget microcontroller

Laboratories ranging the scientific disciplines employ feedback control to regulate variables within their experiments, from the flow of liquids within a microfluidic device to the temperature within a cell incubator. We have built an inexpensive, yet fast and rapidly deployed, feedback control system that is straightforward and flexible to implement from a commercially available Arduino Due microcontroller. This is in comparison with the complex, time-consuming and often expensive electronics that are commonly implemented. As an example of its utility, we apply our feedback controller to the task of stabilizing the main trapping laser of an optical tweezers. The feedback controller, which is inexpensive yet fast and rapidly deployed, was implemented from hacking an open source Arduino Due microcontroller. Our microcontroller based feedback system can stabilize the laser intensity to a few tenths of a per cent at 200 kHz, which is an order of magnitude better than the laser's base specifications, illustrating the utility of these devices. (paper)

10. Dynamic measurements and simulations of airborne picolitre-droplet coalescence in holographic optical tweezers

Bzdek, Bryan R.; Collard, Liam; Sprittles, James E.; Hudson, Andrew J.; Reid, Jonathan P.

2016-08-01

We report studies of the coalescence of pairs of picolitre aerosol droplets manipulated with holographic optical tweezers, probing the shape relaxation dynamics following coalescence by simultaneously monitoring the intensity of elastic backscattered light (EBL) from the trapping laser beam (time resolution on the order of 100 ns) while recording high frame rate camera images (time resolution inhibits quantitative determination of the relaxation time scale. Droplet coalescence was simulated using a computational framework to validate both experimental approaches. The results indicate that time-dependent monitoring of droplet shape from the EBL intensity allows for robust determination of properties such as surface tension and viscosity. Finally, the potential of high frame rate imaging to examine the coalescence of dissimilar viscosity droplets is discussed.

11. In Situ Raman Spectroscopy of COOH-Functionalized SWCNTs Trapped with Optoelectronic Tweezers

Peter J. Pauzauskie

2012-01-01

Full Text Available Optoelectronic tweezers (OETs were used to trap and deposit aqueous dispersions of carboxylic-acid-functionalized single-walled carbon nanotube bundles. Dark-field video microscopy was used to visualize the dynamics of the bundles both with and without virtual electrodes, showing rapid accumulation of carbon nanotubes when optical virtual electrodes are actuated. Raman microscopy was used to probe SWCNT materials following deposition onto metallic fiducial markers as well as during trapping. The local carbon nanotube concentration was observed to increase rapidly during trapping by more than an order of magnitude in less than one second due to localized optical dielectrophoresis forces. This combination of enrichment and spectroscopy with a single laser spot suggests a broad range of applications in physical, chemical, and biological sciences.

12. Mechanism of termination of bacteriophage DNA packaging investigated with optical tweezers

delToro, Damian J.; Smith, Douglas E.

2012-10-01

The genomes of many dsDNA viruses are replicated by a mechanism that produces a long concatemer of multiple genomes. These viruses utilize multifunctional molecular motor complexes referred to as "terminases" that can excise a unit genome length of DNA and package it into preformed viral shells. Remarkably, the terminase motor can initiate packaging at the appropriate start point, translocate DNA, sense when a sufficient length has been packaged, and then switch into a mode where it arrests and cleaves the DNA to release a filled virus particle. We have recently developed an improved method to measure single phage lambda DNA packaging using dual-trap optical tweezers and pre-stalled motor-DNA-procapsid complexes. We are applying this method to test proposed mechanisms for the sensor that triggers termination; specifically a velocity-monitor model vs. energy-monitor model vs. capsid-filling monitor model.

13. Custom-Built Optical Tweezers for Locally Probing the Viscoelastic Properties of Cancer Cells

Tavano, Federica; Bonin, Serena; Pinato, Giulietta; Stanta, Giorgio; Cojoc, Dan

2011-07-01

We report a home built optical tweezers setup to investigate the mechanism of the membrane tether formation from single cells in vitro. Using an optically trapped microbead as probe, we have determined the force-elongation curve during tether formation and extracted several parameters characterizing the viscoelastic behavior of the cell membrane: tether stiffness, force, and viscosity. Breast cancer MDA-MB-231 cells have been studied in two different conditions, at room and physiological temperatures, showing a strong temperature dependence of the visoelastic properties of the cell membrane. To get detailed inside information about the tether formation mechanism we have extended the analysis of the force-elongation curves fitting them with a Kelvin model. These preliminary results are part of a larger project of whose goal is to compare the viscoelastic properties of several types of cancer cell lines, characterized by different aggressiveness and metastatic potential.

14. Characterization of the mechanical properties of HL-1 cardiomyocytes with high throughput magnetic tweezers

We characterized the mechanical properties of cardiomyocyte-like HL-1 cells using our recently developed multi-pole magnetic tweezers. With the optimized design, both high force and high throughput are achieved at the same time. Force up to 100 pN can be applied on a 1 μm diameter superparamagnetic bead in a workspace with 60 μm radius, which is encircled symmetrically by 3 sharp magnetic tips. By adjusting the coil currents, both the strength and direction of force can be controlled. The result shows that both viscosity and shear elastic modulus of HL-1 cells exhibit an approximately log-normal distribution. The cells became stiffer as they matured, consistent with a transition from proliferating cells to contractile muscle tissue. Moreover, the mechanical properties of HL-1 cells show high heterogeneity, which agrees well with their physiological structure

15. Parallel analysis of individual biological cells using multifocal laser tweezers Raman spectroscopy.

Liu, Rui; Taylor, Douglas S; Matthews, Dennis L; Chan, James W

2010-11-01

We report on the development and characterization of a multifocal laser tweezers Raman spectroscopy (M-LTRS) technique for parallel Raman spectral acquisition of individual biological cells. Using a 785-nm diode laser and a time-sharing laser trapping scheme, multiple laser foci are generated to optically trap single polystyrene beads and suspension cells in a linear pattern. Raman signals from the trapped objects are simultaneously projected through the slit of a spectrometer and spatially resolved on a charge-coupled device (CCD) detector with minimal signal crosstalk between neighboring cells. By improving the rate of single-cell analysis, M-LTRS is expected to be a valuable method for studying single-cell dynamics of cell populations and for the development of high-throughput Raman based cytometers. PMID:21073802

16. Trapping and rotating nanoparticles using a plasmonic nano-tweezer with an integrated heat sink.

Wang, Kai; Schonbrun, Ethan; Steinvurzel, Paul; Crozier, Kenneth B

2011-01-01

Although optical tweezers based on far-fields have proven highly successful for manipulating objects larger than the wavelength of light, they face difficulties at the nanoscale because of the diffraction-limited focused spot size. This has motivated interest in trapping particles with plasmonic nanostructures, as they enable intense fields confined to sub-wavelength dimensions. A fundamental issue with plasmonics, however, is Ohmic loss, which results in the water, in which the trapping is performed, being heated and to thermal convection. Here we demonstrate the trapping and rotation of nanoparticles using a template-stripped plasmonic nanopillar incorporating a heat sink. Our simulations predict an ~100-fold reduction in heating compared with previous designs. We further demonstrate the stable trapping of polystyrene particles, as small as 110 nm in diameter, which can be rotated around the nanopillar actively, by manual rotation of the incident linear polarization, or passively, using circularly polarized illumination. PMID:21915111

17. Optical tweezers assisted imaging of the Z-ring in Escherichia coli: measuring its radial width

Carmon, G.; Kumar, P.; Feingold, M.

2014-01-01

Using single-beam, oscillating optical tweezers we can trap and rotate rod-shaped bacterial cells with respect to the optical axis. This technique allows imaging fluorescently labeled three-dimensional sub-cellular structures from different, optimized viewpoints. To illustrate our method we measure D, the radial width of the Z-ring in unconstricted Escherichia coli. We use cells that express FtsZ-GFP and have their cytoplasmic membrane stained with FM4-64. In a vertically oriented cell, both the Z-ring and the cytoplasmic membrane images appear as symmetric circular structures that lend themselves to quantitative analysis. We found that D ≅ 100 nm, much larger than expected.

18. Light-Induced Agglomeration and Diffusion of Different Particles with Optical Tweezers

LI Xue-Cong; SUN Xiu-Dong; LIU Hong-Peng; ZHANG Jian-Long

2010-01-01

@@ The dynamic process of light-induced agglomeration of carbon nanotubes(CNTs),C60 and Escherichia coli(E.coli)in aqueous solutions is demonstrated using an optical tweezers system.Based on the results,the diameter of the agglomerated region and the agglomeration rate increase with the increasing laser power.After the saturation-stable period,CNTs diffuse completely,C60 dusters only diffuse partially,and E.coli never diffuses in the agglomeration region.Theoretical analyses show that the molecular polarization and thermal diffusion of particles play crucial roles in the diffusion process.The results indicate the possibility of using light to aggregate and sort nanoparticles.

19. Microfluidic platform combining droplets and magnetic tweezers: application to HER2 expression in cancer diagnosis

Ferraro, Davide; Champ, Jérôme; Teste, Bruno; Serra, Marco; Malaquin, Laurent; Viovy, Jean-Louis; de Cremoux, Patricia; Descroix, Stephanie

2016-05-01

The development of precision medicine, together with the multiplication of targeted therapies and associated molecular biomarkers, call for major progress in genetic analysis methods, allowing increased multiplexing and the implementation of more complex decision trees, without cost increase or loss of robustness. We present a platform combining droplet microfluidics and magnetic tweezers, performing RNA purification, reverse transcription and amplification in a fully automated and programmable way, in droplets of 250nL directly sampled from a microtiter-plate. This platform decreases sample consumption about 100 fold as compared to current robotized platforms and it reduces human manipulations and contamination risk. The platform’s performance was first evaluated on cell lines, showing robust operation on RNA quantities corresponding to less than one cell, and then clinically validated with a cohort of 21 breast cancer samples, for the determination of their HER2 expression status, in a blind comparison with an established routine clinical analysis.

20. Optical nanofibre integrated into an optical tweezers for particle manipulation, in situ fibre probing, and optical binding studies

Gusachenko, Ivan; Frawley, Mary C; Chormaic, Síle Nic

2015-01-01

Precise control of particle positioning is desirable in many optical propulsion and sorting applications. Here, we develop an integrated platform for particle manipulation consisting of a combined optical nanofibre and optical tweezers system. Individual silica microspheres were introduced to the nanofibre at arbitrary points using the optical tweezers, thereby producing pronounced dips in the fibre transmission. We show that such consistent and reversible transmission modulations depend on both particle and fibre diameter, and can be used as a reference point for in situ nanofibre or particle size measurement. Thence, we combine scanning electron microscope (SEM) size measurements with nanofibre transmission data to provide calibration for particle-based fibre assessment. This integrated optical platform provides a method for selective evanescent field manipulation of micron-sized particles and facilitates studies of optical binding and light-particle interaction dynamics.

1. Self-assembly characteristics of a multipolar donor-acceptor-based bis-pyrene integrated molecular tweezer

Deepak Asthana; Geeta Hundal; Pritam Mukhopadhyay

2014-09-01

A modular design of a molecular tweezer is presented that integrates a multipolar D--A [D: Donor, A: Acceptor] scaffold, 1-aminopyrene-based fluorophore units and L-alanine-based linkers. The synthesis of the molecule is based on two-fold aromatic nucleophilic reactions (ArSN) and coupling reactions of the acid and amino functionalities. This molecule crystallizes in a non-centrosymmteric (P21) space group.We present its rich self-assembly characteristics that involves an array of -stacking interactions. In addition, the molecular tweezer within its cleft forms H-bonding with two dimethylformamide molecules. Such multipolar D--A systems containing chiral and fluorophore units are potential candidatesfor a number of electronic and photonic applications.

2. Real-time observation of ultrafast electron injection at graphene–Zn porphyrin interfaces

2015-02-25

We report on the ultrafast interfacial electron transfer ( ET) between zinc( II) porphyrin ( ZnTMPyP) and negatively charged graphene carboxylate ( GC) using state- of- the- art femtosecond laser spectroscopy with broadband capabilities. The steady- state interaction between GC and ZnTMPyP results in a red- shifted absorption spectrum, providing a clear indication for the binding affinity between ZnTMPyP and GC via electrostatic and p- p stacking interactions. Ultrafast transient absorption ( TA) spectra in the absence and presence of three different GC concentrations reveal ( i) the ultrafast formation of singlet excited ZnTMPyP*, which partially relaxes into a long- lived triplet state, and ( ii) ET from the singlet excited ZnTMPyP* to GC, forming ZnTMPyP + and GC , as indicated by a spectral feature at 650- 750 nm, which is attributed to a ZnTMPyP radical cation resulting from the ET process.

3. Lysine-specific molecular tweezers are broad-spectrum inhibitors of assembly and toxicity of amyloid proteins

Sinha, Sharmistha; Lopes, Dahabada H. J.; Du, Zhenming; Pang, Eric S.; Shanmugam, Akila; Lomakin, Aleksey; Talbiersky, Peter; Tennstaedt, Annette; McDaniel, Kirsten; Bakshi, Reena; Kuo, Pei-Yi; Ehrmann, Michael; Benedek, George B.; Loo, Joseph A.; Klärner, Frank-Gerrit

2011-01-01

Amyloidoses are diseases characterized by abnormal protein folding and self-assembly, for which no cure is available. Inhibition or modulation of abnormal protein self-assembly therefore is an attractive strategy for prevention and treatment of amyloidoses. We examined Lys-specific molecular tweezers and discovered a lead compound termed CLR01, which is capable of inhibiting the aggregation and toxicity of multiple amyloidogenic proteins by binding to Lys residues and disrupting hydrophobic a...

4. Blind predictions of DNA and RNA tweezers experiments with force and torque.

Fang-Chieh Chou

2014-08-01

Full Text Available Single-molecule tweezers measurements of double-stranded nucleic acids (dsDNA and dsRNA provide unprecedented opportunities to dissect how these fundamental molecules respond to forces and torques analogous to those applied by topoisomerases, viral capsids, and other biological partners. However, tweezers data are still most commonly interpreted post facto in the framework of simple analytical models. Testing falsifiable predictions of state-of-the-art nucleic acid models would be more illuminating but has not been performed. Here we describe a blind challenge in which numerical predictions of nucleic acid mechanical properties were compared to experimental data obtained recently for dsRNA under applied force and torque. The predictions were enabled by the HelixMC package, first presented in this paper. HelixMC advances crystallography-derived base-pair level models (BPLMs to simulate kilobase-length dsDNAs and dsRNAs under external forces and torques, including their global linking numbers. These calculations recovered the experimental bending persistence length of dsRNA within the error of the simulations and accurately predicted that dsRNA's "spring-like" conformation would give a two-fold decrease of stretch modulus relative to dsDNA. Further blind predictions of helix torsional properties, however, exposed inaccuracies in current BPLM theory, including three-fold discrepancies in torsional persistence length at the high force limit and the incorrect sign of dsRNA link-extension (twist-stretch coupling. Beyond these experiments, HelixMC predicted that 'nucleosome-excluding' poly(A/poly(T is at least two-fold stiffer than random-sequence dsDNA in bending, stretching, and torsional behaviors; Z-DNA to be at least three-fold stiffer than random-sequence dsDNA, with a near-zero link-extension coupling; and non-negligible effects from base pair step correlations. We propose that experimentally testing these predictions should be powerful next

5. Drug-DNA interactions at single molecule level: A view with optical tweezers

Paramanathan, Thayaparan

Studies of small molecule--DNA interactions are essential for developing new drugs for challenging diseases like cancer and HIV. The main idea behind developing these molecules is to target and inhibit the reproduction of the tumor cells and infected cells. We mechanically manipulate single DNA molecule using optical tweezers to investigate two molecules that have complex and multiple binding modes. Mononuclear ruthenium complexes have been extensively studied as a test for rational drug design. Potential drug candidates should have high affinity to DNA and slow dissociation kinetics. To achieve this, motifs of the ruthenium complexes are altered. Our collaborators designed a dumb-bell shaped binuclear ruthenium complex that can only intercalate DNA by threading through its bases. Studying the binding properties of this complex in bulk studies took hours. By mechanically manipulating a single DNA molecule held with optical tweezers, we lower the barrier to thread and make it fast compared to the bulk experiments. Stretching single DNA molecules with different concentration of drug molecules and holding it at a constant force allows the binding to reach equilibrium. By this we can obtain the equilibrium fractional ligand binding and length of DNA at saturated binding. Fitting these results yields quantitative measurements of the binding thermodynamics and kinetics of this complex process. The second complex discussed in this study is Actinomycin D (ActD), a well studied anti-cancer agent that is used as a prototype for developing new generations of drugs. However, the biophysical basis of its activity is still unclear. Because ActD is known to intercalate double stranded DNA (dsDNA), it was assumed to block replication by stabilizing dsDNA in front of the replication fork. However, recent studies have shown that ActD binds with even higher affinity to imperfect duplexes and some sequences of single stranded DNA (ssDNA). We directly measure the on and off rates by

6. Blind predictions of DNA and RNA tweezers experiments with force and torque.

Chou, Fang-Chieh; Lipfert, Jan; Das, Rhiju

2014-08-01

Single-molecule tweezers measurements of double-stranded nucleic acids (dsDNA and dsRNA) provide unprecedented opportunities to dissect how these fundamental molecules respond to forces and torques analogous to those applied by topoisomerases, viral capsids, and other biological partners. However, tweezers data are still most commonly interpreted post facto in the framework of simple analytical models. Testing falsifiable predictions of state-of-the-art nucleic acid models would be more illuminating but has not been performed. Here we describe a blind challenge in which numerical predictions of nucleic acid mechanical properties were compared to experimental data obtained recently for dsRNA under applied force and torque. The predictions were enabled by the HelixMC package, first presented in this paper. HelixMC advances crystallography-derived base-pair level models (BPLMs) to simulate kilobase-length dsDNAs and dsRNAs under external forces and torques, including their global linking numbers. These calculations recovered the experimental bending persistence length of dsRNA within the error of the simulations and accurately predicted that dsRNA's "spring-like" conformation would give a two-fold decrease of stretch modulus relative to dsDNA. Further blind predictions of helix torsional properties, however, exposed inaccuracies in current BPLM theory, including three-fold discrepancies in torsional persistence length at the high force limit and the incorrect sign of dsRNA link-extension (twist-stretch) coupling. Beyond these experiments, HelixMC predicted that 'nucleosome-excluding' poly(A)/poly(T) is at least two-fold stiffer than random-sequence dsDNA in bending, stretching, and torsional behaviors; Z-DNA to be at least three-fold stiffer than random-sequence dsDNA, with a near-zero link-extension coupling; and non-negligible effects from base pair step correlations. We propose that experimentally testing these predictions should be powerful next steps for

7. High-resolution detection of Brownian motion for quantitative optical tweezers experiments.

Grimm, Matthias; Franosch, Thomas; Jeney, Sylvia

2012-08-01

We have developed an in situ method to calibrate optical tweezers experiments and simultaneously measure the size of the trapped particle or the viscosity of the surrounding fluid. The positional fluctuations of the trapped particle are recorded with a high-bandwidth photodetector. We compute the mean-square displacement, as well as the velocity autocorrelation function of the sphere, and compare it to the theory of Brownian motion including hydrodynamic memory effects. A careful measurement and analysis of the time scales characterizing the dynamics of the harmonically bound sphere fluctuating in a viscous medium directly yields all relevant parameters. Finally, we test the method for different optical trap strengths, with different bead sizes and in different fluids, and we find excellent agreement with the values provided by the manufacturers. The proposed approach overcomes the most commonly encountered limitations in precision when analyzing the power spectrum of position fluctuations in the region around the corner frequency. These low frequencies are usually prone to errors due to drift, limitations in the detection, and trap linearity as well as short acquisition times resulting in poor statistics. Furthermore, the strategy can be generalized to Brownian motion in more complex environments, provided the adequate theories are available. PMID:23005790

8. Force unfolding kinetics of RNA using optical tweezers. I. Effects of experimental variables on measured results

Wen, J -D; Li, P T X; Smith, S B; Bustamante, C; Ritort, F; Tinoco, I

2007-01-01

Experimental variables of optical tweezers instrumentation that affect RNA folding/unfolding kinetics were investigated. A model RNA hairpin, P5ab, was attached to two micron-sized beads through hybrid RNA/DNA handles; one bead was trapped by dual-beam lasers and the other was held by a micropipette. Several experimental variables were changed while measuring the unfolding/refolding kinetics, including handle lengths, trap stiffness, and modes of force applied to the molecule. In constant-force mode where the tension applied to the RNA was maintained through feedback control, the measured rate coefficients varied within 40% when the handle lengths were changed by 10 fold (1.1 to 10.2 Kbp); they increased by two- to three-fold when the trap stiffness was lowered to one third (from 0.1 to 0.035 pN/nm). In the passive mode, without feedback control and where the force applied to the RNA varied in response to the end-to-end distance change of the tether, the RNA hopped between a high-force folded-state and a low-...

9. Mechanics of protein-DNA interaction studied with ultra-fast optical tweezers

Monico, Carina; Tempestini, Alessia; Vanzi, Francesco; Pavone, Francesco S.; Capitanio, Marco

2014-05-01

The lac operon is a well known example of gene expression regulation, based on the specific interaction of Lac repressor protein (LacI) with its target DNA sequence (operator). LacI and other DNA-binding proteins bind their specific target sequences with rates higher than allowed by 3D diffusion alone. Generally accepted models predict a combination of free 3D diffusion and 1D sliding along non-specific DNA. We recently developed an ultrafast force-clamp laser trap technique capable of probing molecular interactions with sub-ms temporal resolution, under controlled pN-range forces. With this technique, we tested the interaction of LacI with two different DNA constructs: a construct with two copies of the O1 operator separated by 300 bp and a construct containing the native E.coli operator sequences. Our measurements show at least two classes of LacI-DNA interactions: long (in the tens of s range) and short (tens of ms). Based on position along the DNA sequence, the observed interactions can be interpreted as specific binding to operator sequences (long events) and transient interactions with nonspecific sequences (short events). Moreover, we observe continuous sliding of the protein along DNA, passively driven by the force applied with the optical tweezers.

10. Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers

We investigated the threading and controlled translocation of individual lambda-DNA (λ-DNA) molecules through solid-state nanopores with piconewton force sensitivity, millisecond time resolution and picoampere ionic current sensitivity with a set-up combining quantitative 3D optical tweezers (OT) with electrophysiology. With our virtually interference-free OT set-up the binding of RecA and single peroxiredoxin protein molecules to λ-DNA was quantitatively investigated during dynamic translocation experiments where effective forces and respective ionic currents of the threaded DNA molecule through the nanopore were measured during inward and outward sliding. Membrane voltage-dependent experiments of reversible single protein/DNA translocation scans yield hysteresis-free, asymmetric single-molecule fingerprints in the measured force and conductance signals that can be attributed to the interplay of optical trap and electrostatic nanopore potentials. These experiments allow an exact localization of the bound protein along the DNA strand and open fascinating applications for label-free detection of DNA-binding ligands, where structural and positional binding phenomena can be investigated at a single-molecule level.

11. Cell manipulation tool with combined microwell array and optical tweezers for cell isolation and deposition

Isolation from rare cells and deposition of sorted cells with high accuracy for further study are critical to a wide range of biomedical applications. In the current paper, we report an automated cell manipulation tool with combined optical tweezers and a uniquely designed microwell array, which functions for recognition, isolation, assembly, transportation and deposition of the interesting cells. The microwell array allows the passive hydrodynamic docking of cells, while offering the opportunity to inspect the interesting cell phenotypes with high spatio-temporal resolution based on the flexible image processing technique. In addition, dynamic and parallel cell manipulation in three dimensions can realize the target cell levitation from microwell and pattern assembly with multiple optical traps. Integrated with the programmed motorized stage, the optically levitated and assembled cells can be transported and deposited to the predefined microenvironment, so the tool can facilitate the integration of other on-chip functionalities for further study without removing these isolated cells from the chip. Experiments on human embryonic stem cells and yeast cells are performed to demonstrate the effectiveness of the proposed cell manipulation tool. Besides the application to cell isolation and deposition, three other biological applications with this tool are also presented. (paper)

12. Acoustical tweezers using single spherically focused piston, X-cut, and Gaussian beams.

Mitri, Farid G

2015-10-01

Partial-wave series expansions (PWSEs) satisfying the Helmholtz equation in spherical coordinates are derived for circular spherically focused piston (i.e., apodized by a uniform velocity amplitude normal to its surface), X-cut (i.e., apodized by a velocity amplitude parallel to the axis of wave propagation), and Gaussian (i.e., apodized by a Gaussian distribution of the velocity amplitude) beams. The Rayleigh-Sommerfeld diffraction integral and the addition theorems for the Legendre and spherical wave functions are used to obtain the PWSEs assuming weakly focused beams (with focusing angle α ⩽ 20°) in the Fresnel-Kirchhoff (parabolic) approximation. In contrast with previous analytical models, the derived expressions allow computing the scattering and acoustic radiation force from a sphere of radius a without restriction to either the Rayleigh (a ≪ λ, where λ is the wavelength of the incident radiation) or the ray acoustics (a ≫λ) regimes. The analytical formulations are valid for wavelengths largely exceeding the radius of the focused acoustic radiator, when the viscosity of the surrounding fluid can be neglected, and when the sphere is translated along the axis of wave propagation. Computational results illustrate the analysis with particular emphasis on the sphere's elastic properties and the axial distance to the center of the concave surface, with close connection of the emergence of negative trapping forces. Potential applications are in single-beam acoustical tweezers, acoustic levitation, and particle manipulation. PMID:26470046

13. A new approach to follow a single extracellular vesicle-cell interaction using optical tweezers.

Prada, Ilaria; Amin, Ladan; Furlan, Roberto; Legname, Giuseppe; Verderio, Claudia; Cojoc, Dan

2016-01-01

Extracellular vesicles (EVs) are spherical membrane structures released by most cells. These highly conserved mediators of intercellular communication carry proteins, lipids, and nucleic acids, and transfer these cellular components between cells by different mechanisms, such as endocytosis, macropinocytosis, or fusion. However, the temporal and spatial dynamics of vesicle-cell interactions still remain largely unexplored. Here we used optical tweezers to drive single EVs produced by microglial cells onto the surface of astrocytes or microglia in primary culture. By visualizing single EV-cell contacts, we observed that microglial vesicles displayed different motilities on the surface of astrocytes compared with microglia. After contact, EVs positioned on astrocytes displayed some minor oscillatory motion around the point of adhesion, while vesicles dragged to microglia displayed quite regular directional movement on the plasma membrane. Both the adhesion and motion of vesicles on glial cells were strongly reduced by cloaking phosphatidylserine (PS) residues, which are externalized on the vesicle membrane and act as determinants for vesicle recognition by target cells. These data identify optical manipulation as a powerful tool to monitor in vitro vesicle-cell dynamics with high temporal and spatial resolution and to determine in a quantitative manner the contribution of surface receptors/extracellular protein ligands to the contact. PMID:26757810

14. Proposed triaxial atomic force microscope contact-free tweezers for nanoassembly

Brown, Keith A; Westervelt, Robert M, E-mail: westervelt@seas.harvard.ed [School of Engineering and Applied Science, Harvard University, Cambridge, MA 02138 (United States)

2009-09-23

We propose a triaxial atomic force microscope contact-free tweezer (TACT) for the controlled assembly of nanoparticles suspended in a liquid. The TACT overcomes four major challenges faced in nanoassembly, as follows. (1) The TACT can hold and position a single nanoparticle with spatial accuracy smaller than the nanoparticle size ({approx}5 nm). (2) The nanoparticle is held away from the surface of the TACT by negative dielectrophoresis to prevent van der Waals forces from making it stick to the TACT. (3) The TACT holds nanoparticles in a trap that is size-matched to the particle and surrounded by a repulsive region so that it will only trap a single particle at a time. (4) The trap can hold a semiconductor nanoparticle in water with a trapping energy greater than the thermal energy. For example, a 5 nm radius silicon nanoparticle is held with 10 k{sub B}T at room temperature. We propose methods for using the TACT as a nanoscale pick-and-place tool to assemble semiconductor quantum dots, biological molecules, semiconductor nanowires, and carbon nanotubes.

15. Holographic Raman tweezers controlled by multi-modal natural user interface

Tomori, Zoltán; Keša, Peter; Nikorovič, Matej; Kaňka, Jan; Jákl, Petr; Šerý, Mojmír; Bernatová, Silvie; Valušová, Eva; Antalík, Marián; Zemánek, Pavel

2016-01-01

Holographic optical tweezers provide a contactless way to trap and manipulate several microobjects independently in space using focused laser beams. Although the methods of fast and efficient generation of optical traps are well developed, their user friendly control still lags behind. Even though several attempts have appeared recently to exploit touch tablets, 2D cameras, or Kinect game consoles, they have not yet reached the level of natural human interface. Here we demonstrate a multi-modal ‘natural user interface’ approach that combines finger and gaze tracking with gesture and speech recognition. This allows us to select objects with an operator’s gaze and voice, to trap the objects and control their positions via tracking of finger movement in space and to run semi-automatic procedures such as acquisition of Raman spectra from preselected objects. This approach takes advantage of the power of human processing of images together with smooth control of human fingertips and downscales these skills to control remotely the motion of microobjects at microscale in a natural way for the human operator.

16. Trapping, manipulation and rapid rotation of NBD-C8 fluorescent single microcrystals in optical tweezers

We have built an optical tweezers experiment based on an inverted microscope to trap and manipulate single crystals of micro or sub-micrometer size made from fluorescent molecules of 4-octylamino-7-nitrobenzoxadiazole (NBD-C8). These single crystals have parallelepiped shapes and exhibit birefringence properties evidenced through optical experiments between crossed polarizers in a polarizing microscope. The crystals are uniaxial with their optical axis oriented along their largest dimension. Trapped in the optical trap, the organic micro-crystals are oriented in such a way that their long axis is along the direction of the beam propagation, and their short axis follows the direction of the linear polarization. Therefore, with linearly polarized light, simply rotating the light polarization can orient the crystal. When using circularly or only elliptically polarized light, the crystal can spontaneously rotate and reach rotation speed of several hundreds of turns per second. A surprising result has been observed: when the incident power is growing up, the rotation speed increases to reach a maximum value and then decreases even when the power is still growing up. Moreover, this evolution is irreversible. Different possible explanations can be considered. The development of a 3D control of the crystals by dynamical holography using liquid crystal spatial modulators will be presented and discussed on the basis of the most recent results obtained. (Author)

17. Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers

Sischka, Andy; Spiering, Andre; Khaksar, Maryam; Laxa, Miriam; König, Janine; Dietz, Karl-Josef; Anselmetti, Dario

2010-11-01

We investigated the threading and controlled translocation of individual lambda-DNA (λ-DNA) molecules through solid-state nanopores with piconewton force sensitivity, millisecond time resolution and picoampere ionic current sensitivity with a set-up combining quantitative 3D optical tweezers (OT) with electrophysiology. With our virtually interference-free OT set-up the binding of RecA and single peroxiredoxin protein molecules to λ-DNA was quantitatively investigated during dynamic translocation experiments where effective forces and respective ionic currents of the threaded DNA molecule through the nanopore were measured during inward and outward sliding. Membrane voltage-dependent experiments of reversible single protein/DNA translocation scans yield hysteresis-free, asymmetric single-molecule fingerprints in the measured force and conductance signals that can be attributed to the interplay of optical trap and electrostatic nanopore potentials. These experiments allow an exact localization of the bound protein along the DNA strand and open fascinating applications for label-free detection of DNA-binding ligands, where structural and positional binding phenomena can be investigated at a single-molecule level.

18. Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers

Sischka, Andy; Spiering, Andre; Anselmetti, Dario [Experimental Biophysics and Applied Nanoscience, Faculty of Physics, Bielefeld University, D-33615 Bielefeld (Germany); Khaksar, Maryam [Fachbereich Physik, University of Konstanz, Fach 688, D-78457 Konstanz (Germany); Laxa, Miriam; Koenig, Janine; Dietz, Karl-Josef, E-mail: andy.sischka@physik.uni-bielefeld.d [Biochemistry and Plant Physiology, Faculty of Biology, Bielefeld University, D-33615 Bielefeld (Germany)

2010-11-17

We investigated the threading and controlled translocation of individual lambda-DNA ({lambda}-DNA) molecules through solid-state nanopores with piconewton force sensitivity, millisecond time resolution and picoampere ionic current sensitivity with a set-up combining quantitative 3D optical tweezers (OT) with electrophysiology. With our virtually interference-free OT set-up the binding of RecA and single peroxiredoxin protein molecules to {lambda}-DNA was quantitatively investigated during dynamic translocation experiments where effective forces and respective ionic currents of the threaded DNA molecule through the nanopore were measured during inward and outward sliding. Membrane voltage-dependent experiments of reversible single protein/DNA translocation scans yield hysteresis-free, asymmetric single-molecule fingerprints in the measured force and conductance signals that can be attributed to the interplay of optical trap and electrostatic nanopore potentials. These experiments allow an exact localization of the bound protein along the DNA strand and open fascinating applications for label-free detection of DNA-binding ligands, where structural and positional binding phenomena can be investigated at a single-molecule level.

19. Haptic Manipulation of Microspheres Using Optical Tweezers Under the Guidance of Artificial Force Fields

Bukusoglu, Ibrahim; Kiraz, Alper; Kurt, Adnan

2007-01-01

Using optical tweezers and a haptic device, microspheres having diameters ranging from 3 to 4 um (floating in a fluid solution) are manipulated in order to form patterns of coupled optical microresonators by assembling the spheres via chemical binding. For this purpose, biotin-coated microspheres trapped by a laser beam are steered and chemically attached to an immobilized streptavidin-coated sphere (i.e. anchor sphere) one by one using an XYZ piezo scanner controlled by a haptic device. The positions of all spheres in the scene are detected using a CCD camera and a collision-free path for each manipulated sphere is generated using the potential field approach. The forces acting on the manipulated particle due to the viscosity of the fluid and the artificial potential field are scaled and displayed to the user through the haptic device for better guidance and control during steering. In addition, a virtual fixture is implemented such that the desired angle of approach and strength are achieved during the bind...

20. Effect of neighboring cells on cell stiffness measured by optical tweezers indentation

Yousafzai, Muhammad S.; Coceano, Giovanna; Mariutti, Alberto; Ndoye, Fatou; Amin, Ladan; Niemela, Joseph; Bonin, Serena; Scoles, Giacinto; Cojoc, Dan

2016-05-01

We report on the modification of mechanical properties of breast cancer cells when they get in contact with other neighboring cells of the same type. Optical tweezers vertical indentation was employed to investigate cell mechanics in isolated and contact conditions, by setting up stiffness as a marker. Two human breast cancer cell lines with different aggressiveness [MCF-7 (luminal breast cancer) and MDA-MB-231 (basal-like breast cancer)] and one normal immortalized breast cell line HBL-100 (normal and myoepithelial) were selected. We found that neighboring cells significantly alter cell stiffness: MDA-MB-231 becomes stiffer when in contact, while HBL-100 and MCF-7 exhibit softer character. Cell stiffness was probed at three cellular subregions: central (above nucleus), intermediate (cytoplasm), and near the leading edge. In an isolated condition, all cells showed a significant regional variation in stiffness: higher at the center and fading toward the leading edge. However, the regional variation becomes statistically insignificant when the cells were in contact with other neighboring cells. The proposed approach will contribute to understand the intriguing temporal sequential alterations in cancer cells during interaction with their surrounding microenvironment.

1. Initiation of bacteriophage Φ29 DNA packaging studied by optical tweezers manipulation of single DNA molecules

Rickgauer, John Peter; Fuller, Derek N.; Hu, Bo; Grimes, Shelley; Jardine, Paul J.; Anderson, Dwight L.; Smith, Douglas E.

2006-08-01

A key step in the life cycle of many viruses, including bacteriophages, adenoviruses, and herpesviruses, is the packaging of replicated viral genomes into pre-assembled proheads by the action of ATP-dependent portal motor complexes. Here we present a method that allows the initiation of packaging by single complexes to be studied using optical tweezers. A procedure is developed for assembling phage Φ29 prohead-motor complexes, which are demonstrated to bind and begin translocation of a target DNA molecule within only a few seconds. We show that the Φ29 DNA terminal protein (gene product 3), which functions to prime DNA replication, also has a dramatic effect on packaging. The DNA tether length measured immediately after binding varied from ~30-100% of the full length, yet shortened monotonically, indicating that packaging does not strictly begin at the terminal end of the DNA. Removal of the terminal protein eliminated this variability, causing packaging to initiate at or very near the end of the DNA. These findings, taken together with electron microscopy data, suggest that rather than simply threading into the portal, the motor captures and dynamically tensions a DNA loop, and that the function of the terminal protein is to load DNA segments on both sides of the loop junction onto separate DNA translocating units.

2. A model for inertial particle trapping locations in hydrodynamic tweezers arrays

We present a model for the trapping of particles with finite inertia in the microscale viscous steady streaming flow of hydrodynamic tweezers. Devices containing a square array and an offset array of cylindrical posts of radius 25 µm were fabricated. As water is oscillated at small amplitude (s < 5 µm) and audible frequency (5000 Hz), highly symmetric microeddies form causing the fluid and particles suspended in the fluid to transport through the device. We image the flows by using 0.5 µm radius fluorescent polystyrene particles, and demonstrate trapping with larger 5 µm radius polystyrene particles. The streaming flow fields are simulated numerically using a fast analytic-numeric approach, and inertial particle motion is determined using the well-known Maxey–Riley equation for small Stokes number (St) particle motion. The St-dependent period-averaged particle velocity is used to describe the effects of inertia on particle trapping locations. We find the St-dependence of trapping location depends on the underlying symmetry of the flow. Only traps located near eddy centers are affected by particle inertia. (paper)

3. On chip single-cell separation and immobilization using optical tweezers and thermosensitive hydrogel.

Arai, Fumihito; Ng, Chinaik; Maruyama, Hisataka; Ichikawa, Akihiko; El-Shimy, Haitham; Fukuda, Toshio

2005-12-01

A novel approach appropriate for rapid separation and immobilization of a single cell by concomitantly utilizing laser manipulation and locally thermosensitive hydrogelation is proposed in this paper. We employed a single laser beam as optical tweezers for separating a target cell and locating it adjacent to a fabricated, transparent micro heater. Simultaneously, the target cell is immobilized or partially entrapped by heating the thermosensitive hydrogel with the micro heater. The state of the thermosensitive hydrogel can be switched from sol to gel and gel to sol by controlling the temperature through heating and cooling by the micro heater. After other unwanted cells are removed by the high-speed cleaning flow in the microchannel, the entrapped cell is successfully isolated. It is possible to collect the immobilized target cell for analysis or culture by switching off the micro heater and releasing the cell from the entrapment. We demonstrated that the proposed approach is feasible for rapid manipulation, immobilization, cleaning, isolation and extraction of a single cell. The experimental results are shown here. PMID:16286972

4. Uncoiling mechanism of Klebsiella pneumoniae type 3 pili measured by using optical tweezers

Chen, Feng-Jung; Chan, Chia-Han; Liu, Kuo-Liang; Huang, Ying-Jung; Peng, Hwei-Ling; Chang, Hwan-You; Yew, Tri-Rung; Hsu, Ken Y.; Hsu, Long

2007-09-01

Pili are bacterial appendages that play many important roles in bacterial behaviors, physiology and interaction with hosts. Via pili, bacteria are able to adhere to, migrate onto, and colonize on host cells, mechanically. Different from the most studied type 1 and P type pili, which are rigid and thick with an average of 6~7 nm in diameter, type 3 pili are relatively tiny (3-5 nm in diameter) and flexible, and their biophysical properties remains unclear. By using optical tweezers, we found that the elongation processes of type 3 pili are divided into three phases: (1) elastic elongation, (2) uncoiling elongation, and (3) intrinsic elongation, separately. Besides, the uncoiling force of the recombinant pili displayed on the surface of E. coli [pmrkABCD V1F] is measured 20 pN in average stronger than that of E. coli [pmrkABCD V1]. This suggests that pilin MrkF is involved in determining the mechanical properties of the type 3 pili.

5. Single-molecule kinetics under force: probing protein folding and enzymatic activity with optical tweezers

Wong, Wesley

2010-03-01

Weak non-covalent bonds between and within single molecules govern many aspects of biological structure and function (e.g. DNA base-paring, receptor-ligand binding, protein folding, etc.) In living systems, these interactions are often subject to mechanical forces, which can greatly alter their kinetics and activity. My group develops and applies novel single-molecule manipulation techniques to explore and quantify these force-dependent kinetics. Using optical tweezers, we have quantified the force-dependent unfolding and refolding kinetics of different proteins, including the cytoskeletal protein spectrin in collaboration with E. Evans's group [1], and the A2 domain of the von Willebrand factor blood clotting protein in collaboration with T. Springer's group [2]. Furthermore, we have studied the kinetics of the ADAMTS13 enzyme acting on a single A2 domain, and have shown that physiolgical forces in the circulation can act as a cofactor for enzymatic cleavage, regulating hemostatic activity [2]. References: 1. E. Evans, K. Halvorsen, K. Kinoshita, and W.P. Wong, Handbook of Single Molecule Biophysics, P. Hinterdorfer, ed., Springer (2009). 2. X. Zhang, K. Halvorsen, C.-Z. Zhang, W.P. Wong, and T.A. Springer, Science 324 (5932), 1330-1334 (2009).

6. Probing Protein Folding Kinetics with High-resolution, Stabilized Optical Tweezers

Wong, Wesley; Halvorsen, Ken

2009-03-01

Single-molecule techniques provide a powerful means of exploring molecular transitions such as the unfolding and refolding of a protein. However, the quantification of bi-directional transitions and near-equilibrium phenomena poses unique challenges, and is often limited by the detection resolution and long-term stability of the instrument. We have developed unique optical tweezers methods that address these problems, including an interference-based method for high-resolution 3D bead tracking (˜1 nm laterally, ˜0.3 nm vertically, at > 100 Hz), and a continuous autofocus system that stabilizes the trap height to within 1-2 nm longterm [1,2]. We have used our instruments to quantify the force-dependent unfolding and refolding kinetics of single protein domains (e.g. spectrin in collaboration with E. Evans). These single-molecule studies are presented, together with the accompanying probabilistic analysis that we have developed. References: 1. W.P. Wong, V. Heinrich, E. Evans, Mat. Res. Soc. Symp. Proc., 790, P5.1-P5.10 (2004). 2. V. Heinrich, W.P. Wong, K. Halvorsen, E. Evans, Langmuir, 24, 1194-1203 (2008).

7. Optical tweezers reveal a dynamic mechanical response of cationic peptide-DNA complexes

Lee, Amy; Zheng, Tai; Sucayan, Sarah; Chou, Szu-Ting; Tricoli, Lucas; Hustedt, Jason; Kahn, Jason; Mixson, A. James; Seog, Joonil

2013-03-01

Nonviral carriers have been developed to deliver nucleic acids by forming nanoscale complexes; however, there has been limited success in achieving high transfection efficiency. Our hypothesis is that a factor affecting gene delivery efficiency is the mechanical response of the condensed complex. To begin to test this hypothesis, we directly measured the mechanical properties of DNA-carrier complexes using optical tweezers. Histidine-lysine (HK) polymer, Asparagine-lysine (NK) polymer and poly-L-lysine were used to form complexes with a single DNA molecule. As carriers were introduced, a sudden decrease in DNA extension occurrs at a force level which is defined as critical force (Fc). Fc is carrier and concentration dependent. Pulling revealed reduction in DNA extension length for HK-DNA complexes. The characteristics of force profiles vary by agent and can be dynamically manipulated by changes in environmental conditions such as ionic strength of the buffer as well as pH. Heparin can remove cationic reagents which are otherwise irreversibly bound to DNA. The implications for optimizing molecular interactions to enhance transfection efficiency will be discussed.

8. Stereoscopic particle tracking for 3D touch, vision and closed-loop control in optical tweezers

Force measurement in an interactive 3D micromanipulation system can allow the user to make delicate adjustments, and to explore surfaces with touch as well as vision. We present a system to achieve this on the micron scale using stereoscopic particle tracking combined with holographic optical tweezers, which can track particles with nanometre accuracy. 2D tracking of particles in each of the stereo images gives 3D positions for each particle. This takes less than 200 µs per image pair, using a 1D 'symmetry transform' applied to each row and column of a 2D image, which can maintain tracking of particles throughout the 10 µm axial range. The only parameters required are the geometry of the imaging system, and therefore there is no need to recalibrate for different particle sizes or refractive indices. Consequently, we can calculate the force exerted by the optical trap in real time at 1 kilohertz, allowing us to implement a force-feedback interface (with a loop rate of 400 Hz). In combination with our OpenGL hologram calculation engine, the system has a closed-loop bandwidth of 20 Hz. This allows us to stabilize trapped particles axially through active feedback, cancelling out some Brownian motion. For the weak traps we use here (spring constant k≈2 pN µm−1), this results in a threefold increase in axial stiffness. We demonstrate the 3D interface by probing an oil droplet, mapping out its surface in the y–z plane

9. Determination of femto Newton forces and fluid viscosity using optical tweezers: application to Leishmania amazonensis

Fontes, Adriana; Giorgio, Selma; de Castro, Archimedes B., Jr.; Neto, Vivaldo M.; Pozzo, Liliana d. Y.; Marques, Gustavo P.; Barbosa, Luiz C.; Cesar, Carlos L.

2005-03-01

The objective of this research is to use the displacements of a polystyrene microsphere trapped by an optical tweezers (OT) as a force transducer in mechanical measurements in life sciences. To do this we compared the theoretical optical and hydrodynamic models with experimental data under a broad variation of parameters such as fluid viscosity, refractive index, drag velocity and wall proximities. The laser power was measured after the objective with an integration sphere because normal power meters do not provide an accurate measurement for beam with high numerical apertures. With this careful laser power determination the plot of the optical force (calculated by the particle displacement) versus hydrodynamic force (calculated by the drag velocity) under very different conditions shows an almost 45 degrees straight line. This means that hydrodynamic models can be used to calibrate optical forces and vice-versa. With this calibration we observed the forces of polystyrene bead attached to the protozoa Leishmania amazonensis, responsible for a serious tropical disease. The force range is from 200 femto Newtons to 4 pico Newtons and these experiments shows that OT can be used for infection mechanism and chemotaxis studies in parasites. The other application was to use the optical force to measure viscosities of few microliters sample. Our result shows 5% accuracy measurements.

10. Space-time-wavelength mapping: a new approach for electronic control of optical tweezers

Rahman, Shah; Zhao, Qiancheng; Atasever, Tuva; Boyraz, Ozdal

2015-01-01

We present a new approach for electronic control of optical tweezers. The key technique, called 'space-time-wavelength mapping', involves time-domain modulation which is translated onto spatial domain by diffraction and enables direct control of location and polarity of force hot-spots created by Lorentz force (gradient force). In this study 150 fs optical pulses are dispersed in time and space to achieve a focused elliptical beam that is ~20 {\\mu}m long and ~2 {\\mu}m wide. In order to manipulate the intensity gradient along the beam at the focal spot, we use an electro-optic modulator to modulate power spectral distribution of the femtosecond beam after temporal dispersion. The electro-optic modulator is supplied with a chosen RF waveform that dictates the manipulation of the power spectral distribution. By choosing the appropriate RF waveform, it is possible to create force fields for cell stretching and compression as well as multiple hot spots (of > 200 pN force) for attractive or repulsive forces. We pre...

11. Force unfolding kinetics of RNA using optical tweezers. II. Modeling experiments

Manosas, M; Li, P T X; Smith, S B; Bustamante, C; Tinoco, I; Ritort, F

2007-01-01

By exerting mechanical force it is possible to unfold/refold RNA molecules one at a time. In a small range of forces, an RNA molecule can hop between the folded and the unfolded state with force-dependent kinetic rates. Here, we introduce a mesoscopic model to analyze the hopping kinetics of RNA hairpins in an optical tweezers setup. The model includes different elements of the experimental setup (beads, handles and RNA sequence) and limitations of the instrument (time lag of the force-feedback mechanism and finite bandwidth of data acquisition). We investigated the influence of the instrument on the measured hopping rates. Results from the model are in good agreement with the experiments reported in the companion article (1). The comparison between theory and experiments allowed us to infer the values of the intrinsic molecular rates of the RNA hairpin alone and to search for the optimal experimental conditions to do the measurements. We conclude that long handles and soft laser traps represent the best cond...

12. Optical tweezers based measurement of PLGA-NP interaction with prostate cancer cells

Blesener, Thea; Mondal, Argha; Menon, Jyothi U.; Nguyen, Kytai T.; Mohanty, Samarendra

2013-02-01

In order to quantify the binding capacities of polymeric, biodegradable and biocompatible poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs), conjugated with either R11 peptides or Folic Acid, the strength by detach from prostate cancer cells (PCCs) was measured via optical tweezers based measurements. Specific nanoparticle drug delivery eliminates the previously used diffuse, full-body application of potent cancer drugs by localizing drug delivery to malignant cells. Precise monitoring of NP position in the trap near the PCC membrane using a fluorescence imaging based method enabled calibration of the trap stiffness and subsequent force measurements. By defining the force with which the many diverse conjugates and coatings of different types of NPs bind the vast array of cancer cell types, chemotherapeutic drugs can be delivered in a specific manner with the optimal particle and corresponding conjugates. Further, and most significantly, the rupture force measurements will reveal whether or not targeted nanoparticles can overcome the force of blood attempting to pull the particle from designated cells. Our preliminary study revealed that the binding between PLGA-NPs and prostate cancer cells is enhanced by coating with folic acid or R11 peptides. These conjugates increase the force required to detach the particle thus allowing particles to overcome drag force of the blood in prostate capillary systems.

13. Studying red blood cell agglutination by measuring membrane viscosity with optical tweezers

Fernandes, Heloise P.; Fontes, Adriana; de Thomaz, André A.; Barbosa, Luiz C.; Barjas-Castro, Maria L.; Cesar, Carlos L.

2007-09-01

The red blood cell (RBC) viscoelastic membrane contains proteins and glycoproteins embedded in a fluid lipid bilayer that are responsible for cell agglutination. Manipulating RBCs rouleaux with a double optical tweezers, we observed that the cells slide easily one over the others but are strongly connected by their edges. An explanation for this behavior could be the fact that when the cells slide one over the others, proteins are dragged through the membrane. It confers to the movement a viscous characteristic that is dependent of the velocity between the RBCs and justifies why is so easy to slide them apart. Therefore, in a first step of this work, by measuring the force as a function of the relative velocity between two cells, we confirmed this assumption and used this viscous characteristic of the RBC rouleaux to determine the apparent membrane viscosity of the cell. As this behavior is related to the proteins interactions, we can use the apparent membrane viscosity to obtain a better understanding about cell agglutination. Methods related to cell agglutination induced by antigen-antibody interactions are the basis of most of tests used in transfusion centers. Then, in a second step of this work, we measured the apparent membrane viscosity using antibodies. We observed that this methodology is sensitive to different kinds of bindings between RBCs. Better comprehension of the forces and bindings between RBCs could improve the sensibility and specificity of the hemagglutination reactions and also guides the development of new potentiator substances.

14. Power spectrum analysis for optical tweezers. II: Laser wavelength dependence of parasitic filtering, and how to achieve high bandwidth

Berg-Sørensen, Kirstine; Peterman, Erwin J G; Weber, Tom; Schmidt, Christoph F; Flyvbjerg, Henrik

2006-01-01

the high transparency of silicon to near-infrared light. A simple model that accounts for this phenomenon is here solved for frequencies up to 100 kHz and for laser wavelengths between 750 and 1064 nm. The solution is applied to experimental data in the same range, and is demonstrated to give this......In a typical optical tweezers detection system, the position of a trapped object is determined from laser light impinging on a quadrant photodiode. When the laser is infrared and the photodiode is of silicon, they can act together as an unintended low-pass filter. This parasicit effect is due to...

15. Direct measurement of the dielectrophoresis forces acting on micro-objects using optical tweezers and a simple microfluidic chip

Park, In Soo; Park, Se Hee; Yoon, Dae Sung; Lee, Sang Woo; Kim, Beop-Min

2014-09-01

We constructed a reliable frequency-dependent dielectrophoretic (DEP) force measurement system based on optical tweezers and a microfluidic chip. Using this system, we directly measured the frequency-dependent DEP forces acting on polystyrene beads while varying various parameters, which were all verified by theoretical simulations. We also investigated the DEP characteristics of non-functionalized and carboxyl-functionalized polystyrene beads in solutions with different conductivities by associating the measured crossover frequencies with a theoretical DEP model. This system can be used as a quantifying tool for surface conductance assays by characterizing the DEP forces acting on micro-objects in various experimental conditions.

16. Safety and pharmacological characterization of the molecular tweezer CLR01 - a broad-spectrum inhibitor of amyloid proteins' toxicity.

Attar, A.; Chan, WTC; Klärner, FG; Schrader, T.; Bitan, G.

2014-01-01

The "molecular tweezer" CLR01 is a broad-spectrum inhibitor of abnormal protein self-assembly, which acts by binding selectively to Lys residues. CLR01 has been tested in several in vitro and in vivo models of amyloidoses all without signs of toxicity. With the goal of developing CLR01 as a therapeutic drug for Alzheimer's disease and other amyloidoses, here we studied its safety and pharmacokinetics. Toxicity studies were performed in 2-m old wild-type mice. Toxicity was evaluated by serum c...

17. Guest-responsive structural adaptation of a rationally-designed molecular tweezer based on Tröger’s base

Ishita Neogi; Alankriti Bajpai; Jarugu Narasimha Moorthy

2014-09-01

We have designed and synthesized a modified Tröger’s base TB in which the sterically-rigidified aryl rings that protrude into its groove were envisaged to preclude self-inclusion. From a limited preliminary experimentation, TB has been found to exhibit guest inclusion. The X-ray determined structures of the crystals of guest-free TB and its inclusion compounds with acetonitrile and -dichlorobenzene reveal remarkable adaptability of the TB core to undergo subtle structural changes in response to the guest that is included. The structural analyses demonstrate the fact that TB behaves like a molecular tweezer.

18. Hydrogel-colloid interfacial interactions: a study of tailored adhesion using optical tweezers.

Sheikhi, Amir; Hill, Reghan J

2016-08-21

Dynamics of colloidal particles adhering to soft, deformable substrates, such as tissues, biofilms, and hydrogels play a key role in many biological and biomimetic processes. These processes, including, but not limited to colloid-based delivery, stitching, and sorting, involve microspheres exploring the vicinity of soft, sticky materials in which the colloidal dynamics are affected by the fluid environment (e.g., viscous coupling), inter-molecular interactions between the colloids and substrates (e.g., Derjaguin-Landau-Verwey-Overbeek (DLVO) theory), and the viscoelastic properties of contact region. To better understand colloidal dynamics at soft interfaces, an optical tweezers back-focal-plane interferometry apparatus was developed to register the transverse Brownian motion of a silica microsphere in the vicinity of polyacrylamide (PA) hydrogel films. The time-dependent mean-squared displacements are well described by a single exponential relaxation, furnishing measures of the transverse interfacial diffusion coefficient and binding stiffness. Substrates with different elasticities were prepared by changing the PA crosslinking density, and the inter-molecular interactions were adjusted by coating the microspheres with fluid membranes. Stiffer PA hydrogels (with bulk Young's moduli ≈1-10 kPa) immobilize the microspheres more firmly (lower diffusion coefficient and position variance), and coating the particles with zwitterionic lipid bilayers (DOPC) completely eliminates adhesion, possibly by repulsive dispersion forces. Remarkably, embedding polyethylene glycol-grafted lipid bilayers (DSPE-PEG2k-Amine) in the zwitterionic fluid membranes produces stronger adhesion, possibly because of polymer-hydrogel attraction and entanglement. This study provides new insights to guide the design of nanoparticles and substrates with tunable adhesion, leading to smarter delivery, sorting, and screening of micro- and nano-systems. PMID:27425660

19. Investigation into local cell mechanics by atomic force microscopy mapping and optical tweezer vertical indentation.

Coceano, G; Yousafzai, M S; Ma, W; Ndoye, F; Venturelli, L; Hussain, I; Bonin, S; Niemela, J; Scoles, G; Cojoc, D; Ferrari, E

2016-02-12

Investigating the mechanical properties of cells could reveal a potential source of label-free markers of cancer progression, based on measurable viscoelastic parameters. The Young's modulus has proved to be the most thoroughly studied so far, however, even for the same cell type, the elastic modulus reported in different studies spans a wide range of values, mainly due to the application of different experimental conditions. This complicates the reliable use of elasticity for the mechanical phenotyping of cells. Here we combine two complementary techniques, atomic force microscopy (AFM) and optical tweezer microscopy (OTM), providing a comprehensive mechanical comparison of three human breast cell lines: normal myoepithelial (HBL-100), luminal breast cancer (MCF-7) and basal breast cancer (MDA-MB-231) cells. The elastic modulus was measured locally by AFM and OTM on single cells, using similar indentation approaches but different measurement parameters. Peak force tapping AFM was employed at nanonewton forces and high loading rates to draw a viscoelastic map of each cell and the results indicated that the region on top of the nucleus provided the most meaningful results. OTM was employed at those locations at piconewton forces and low loading rates, to measure the elastic modulus in a real elastic regime and rule out the contribution of viscous forces typical of AFM. When measured by either AFM or OTM, the cell lines' elasticity trend was similar for the aggressive MDA-MB-231 cells, which were found to be significantly softer than the other two cell types in both measurements. However, when comparing HBL-100 and MCF-7 cells, we found significant differences only when using OTM. PMID:26683826

20. Investigation into local cell mechanics by atomic force microscopy mapping and optical tweezer vertical indentation

Coceano, G.; Yousafzai, M. S.; Ma, W.; Ndoye, F.; Venturelli, L.; Hussain, I.; Bonin, S.; Niemela, J.; Scoles, G.; Cojoc, D.; Ferrari, E.

2016-02-01

Investigating the mechanical properties of cells could reveal a potential source of label-free markers of cancer progression, based on measurable viscoelastic parameters. The Young’s modulus has proved to be the most thoroughly studied so far, however, even for the same cell type, the elastic modulus reported in different studies spans a wide range of values, mainly due to the application of different experimental conditions. This complicates the reliable use of elasticity for the mechanical phenotyping of cells. Here we combine two complementary techniques, atomic force microscopy (AFM) and optical tweezer microscopy (OTM), providing a comprehensive mechanical comparison of three human breast cell lines: normal myoepithelial (HBL-100), luminal breast cancer (MCF-7) and basal breast cancer (MDA-MB-231) cells. The elastic modulus was measured locally by AFM and OTM on single cells, using similar indentation approaches but different measurement parameters. Peak force tapping AFM was employed at nanonewton forces and high loading rates to draw a viscoelastic map of each cell and the results indicated that the region on top of the nucleus provided the most meaningful results. OTM was employed at those locations at piconewton forces and low loading rates, to measure the elastic modulus in a real elastic regime and rule out the contribution of viscous forces typical of AFM. When measured by either AFM or OTM, the cell lines’ elasticity trend was similar for the aggressive MDA-MB-231 cells, which were found to be significantly softer than the other two cell types in both measurements. However, when comparing HBL-100 and MCF-7 cells, we found significant differences only when using OTM.

1. Optical tweezers for single molecule force spectroscopy on bacterial adhesion organelles

Andersson, Magnus; Axner, Ove; Uhlin, Bernt Eric; Fällman, Erik

2006-08-01

Instrumentation and methodologies for single molecule force spectroscopy on bacterial adhesion organelles by the use of force measuring optical tweezers have been developed. A thorough study of the biomechanical properties of fimbrial adhesion organelles expressed by uropathogenic E. coli, so-called pili, is presented. Steady-state as well as dynamic force measurements on P pili, expressed by E. coli causing pyelonephritis, have revealed, among other things, various unfolding and refolding properties of the helical structure of P pili, the PapA rod. Based on these properties an energy landscape model has been constructed by which specific biophysical properties of the PapA rod have been extracted, e.g. the number of subunits, the length of a single pilus, bond lengths and activation energies for bond opening and closure. Moreover, long time repetitive measurements have shown that the rod can be unfolded and refolded repetitive times without losing its intrinsic properties. These properties are believed to be of importance for the bacteria's ability to maintain close contact with host cells during initial infections. The results presented are considered to be of importance for the field of biopolymers in general and the development of new pharmaceuticals towards urinary tract infections in particular. The results show furthermore that the methodology can be used to gain knowledge of the intrinsic biomechanical function of adhesion organelles. The instrumentation is currently used for characterization of type 1 pili, expressed by E. coli causing cystitis, i.e. infections in the bladder. The first force spectrometry investigations of these pili will be presented.

2. Annular spherically focused ring transducers for improved single-beam acoustical tweezers

Mitri, F. G.

2016-02-01

The use of ultrasonic transducers with a central hollow is suggested for improved single-beam acoustical tweezers applications. Within the framework of the Fresnel-Kirchhoff parabolic approximation, a closed-form partial-wave series expansion (PWSE) for the incident velocity potential (or pressure) field is derived for an annular spherically focused ring (asfr) with uniform vibration across its surface in spherical coordinates. The Rayleigh-Sommerfeld diffraction integral and the addition theorems for the Legendre and spherical wave functions are used to obtain the PWSE assuming a weakly focused beam (with a focusing angle α ≤ 20°). The PWSE allows evaluating the incident field from the finite asfr in 3D. Moreover, the obtained solution allows computing efficiently the acoustic scattering and radiation force on a sphere centered on the beam's axis of wave propagation. The analytical solution is valid for wavelengths largely exceeding the radius of the asfr and when the viscosity of the surrounding fluid can be neglected. Numerical predictions for the beam-forming, scattering, and axial time-averaged radiation force are performed with particular emphasis on the asfr thickness, the axial distance separating the sphere from the center of the transducer, the (non-dimensional) size of the transducer, as well as the sphere's elastic properties without restriction to the long- (i.e., Rayleigh) or the short-wavelength (i.e., ray acoustics) regimes. Potential applications of the present solution are in beam-forming design, particle tweezing, and manipulation due to negative forces using ultrasonic asfr transducers.

3. Annular spherically focused ring transducers for improved single-beam acoustical tweezers

Mitri, F. G., E-mail: F.G.Mitri@ieee.org [Chevron, Area 52 Technology—ETC, Santa Fe, New Mexico 87508 (United States)

2016-02-14

The use of ultrasonic transducers with a central hollow is suggested for improved single-beam acoustical tweezers applications. Within the framework of the Fresnel-Kirchhoff parabolic approximation, a closed-form partial-wave series expansion (PWSE) for the incident velocity potential (or pressure) field is derived for an annular spherically focused ring (asfr) with uniform vibration across its surface in spherical coordinates. The Rayleigh-Sommerfeld diffraction integral and the addition theorems for the Legendre and spherical wave functions are used to obtain the PWSE assuming a weakly focused beam (with a focusing angle α ≤ 20°). The PWSE allows evaluating the incident field from the finite asfr in 3D. Moreover, the obtained solution allows computing efficiently the acoustic scattering and radiation force on a sphere centered on the beam's axis of wave propagation. The analytical solution is valid for wavelengths largely exceeding the radius of the asfr and when the viscosity of the surrounding fluid can be neglected. Numerical predictions for the beam-forming, scattering, and axial time-averaged radiation force are performed with particular emphasis on the asfr thickness, the axial distance separating the sphere from the center of the transducer, the (non-dimensional) size of the transducer, as well as the sphere's elastic properties without restriction to the long- (i.e., Rayleigh) or the short-wavelength (i.e., ray acoustics) regimes. Potential applications of the present solution are in beam-forming design, particle tweezing, and manipulation due to negative forces using ultrasonic asfr transducers.

4. Single molecule measurements of DNA helicase activity with magnetic tweezers and t-test based step-finding analysis.

Seol, Yeonee; Strub, Marie-Paule; Neuman, Keir C

2016-08-01

Magnetic tweezers is a versatile and easy to implement single-molecule technique that has become increasingly prevalent in the study of nucleic acid based molecular motors. Here, we provide a description of the magnetic tweezers instrument and guidelines for measuring and analyzing DNA helicase activity. Along with experimental methods, we describe a robust method of single-molecule trajectory analysis based on the Student's t-test that accommodates continuous transitions in addition to the discrete transitions assumed in most widely employed analysis routines. To illustrate the single-molecule unwinding assay and the analysis routine, we provide DNA unwinding measurements of Escherichia coli RecQ helicase under a variety of conditions (Na+, ATP, temperature, and DNA substrate geometry). These examples reveal that DNA unwinding measurements under various conditions can aid in elucidating the unwinding mechanism of DNA helicase but also emphasize that environmental effects on DNA helicase activity must be considered in relation to in vivo activity and mechanism. PMID:27131595

5. Self-Locking Optoelectronic Tweezers for Single-Cell and Microparticle Manipulation across a Large Area in High Conductivity Media

Yang, Yajia; Mao, Yufei; Shin, Kyeong-Sik; Chui, Chi On; Chiou, Pei-Yu

2016-03-01

Optoelectronic tweezers (OET) has advanced within the past decade to become a promising tool for cell and microparticle manipulation. Its incompatibility with high conductivity media and limited throughput remain two major technical challenges. Here a novel manipulation concept and corresponding platform called Self-Locking Optoelectronic Tweezers (SLOT) are proposed and demonstrated to tackle these challenges concurrently. The SLOT platform comprises a periodic array of optically tunable phototransistor traps above which randomly dispersed single cells and microparticles are self-aligned to and retained without light illumination. Light beam illumination on a phototransistor turns off the trap and releases the trapped cell, which is then transported downstream via a background flow. The cell trapping and releasing functions in SLOT are decoupled, which is a unique feature that enables SLOT’s stepper-mode function to overcome the small field-of-view issue that all prior OET technologies encountered in manipulation with single-cell resolution across a large area. Massively parallel trapping of more than 100,000 microparticles has been demonstrated in high conductivity media. Even larger scale trapping and manipulation can be achieved by linearly scaling up the number of phototransistors and device area. Cells after manipulation on the SLOT platform maintain high cell viability and normal multi-day divisibility.

6. Axial and transverse acoustic radiation forces on a fluid sphere placed arbitrarily in Bessel beam standing wave tweezers

Mitri, F.G., E-mail: mitri@chevron.com

2014-03-15

The axial and transverse radiation forces on a fluid sphere placed arbitrarily in the acoustical field of Bessel beams of standing waves are evaluated. The three-dimensional components of the time-averaged force are expressed in terms of the beam-shape coefficients of the incident field and the scattering coefficients of the fluid sphere using a partial-wave expansion (PWE) method. Examples are chosen for which the standing wave field is composed of either a zero-order (non-vortex) Bessel beam, or a first-order Bessel vortex beam. It is shown here, that both transverse and axial forces can push or pull the fluid sphere to an equilibrium position depending on the chosen size parameter ka (where k is the wave-number and a the sphere’s radius). The corresponding results are of particular importance in biophysical applications for the design of lab-on-chip devices operating with Bessel beams standing wave tweezers. Moreover, potential investigations in acoustic levitation and related applications in particle rotation in a vortex beam may benefit from the results of this study. -- Highlights: •The axial and transverse forces on a fluid sphere in acoustical Bessel beams tweezers are evaluated. •The attraction or repulsion to an equilibrium position in the standing wave field is examined. •Potential applications are in particle manipulation using standing waves.

7. 3Dtrapping and manipulation of micro-particles using holographic optical tweezers with optimized computer-generated holograms

Tao Tao; Jing Li; Qian Long; Xiaoping Wu

2011-01-01

A multi-plane adaptive-additive algorithm is developed for optimizing computer-generated holograms for the reconstruction of traps in three-dimensional (3D) spaces. This algorithm overcomes the converging stagnation problem of the traditional multi-plane Gerchberg-Saxton algorithm and improves the diffraction efficiency of the holograms effectively. The optimized holograms are applied in a holographic optical tweezers (HOT) platform. Additionally, a computer program is developed and integrated into the HOT platform for the purpose of achieving the interactive control of traps. Experiments demonstrate that the manipulation of micro-particles into the 3D structure with optimized holograms can be carried out effectively on the HOT platform.%A multi-plane adaptive-additive algorithm is developed for optimizing computer-generated holograms for the reconstruction of traps in three-dimensional (3D) spaces.This algorithm overcomes the converging stagnation problem of the traditional multi-plane Gerchberg-Saxton algorithm and improves the diffraction efficiency of the holograms effectively.The optimized holograms are applied in a holographic optical tweezers (HOT) platform.Additionally,a computer program is developed and integrated into the HOT platform for the purpose of achieving the interactive control of traps.Experiments demonstrate that the manipulation of micro-particles into the 3D structure with optimized holograms can be carried out effectively on the HOT platform.

8. Single-molecule manipulation of double-stranded DNA using optical tweezers: Interaction studies of DNA with RecA and YOYO-1

Bennink, Martin L.; Scharer, Orlando D.; Kanaar, Ronald; Sakata-Sogawa, Kumiko; Schins, Juleon M.; Kanger, Johannes S.; Grooth, de Bart G.; Greve, Jan

1999-01-01

By using optical tweezers and a specially designed flow cell with an integrated glass micropipette, we constructed a setup similar to that of Smith et al. (Science 271:795-799, 1996) in which an individual double-stranded DNA (dsDNA) molecule can be captured between two polystyrene beads. The first

9. Power spectrum analysis with least-squares fitting: Amplitude bias and its elimination, with application to optical tweezers and atomic force microscope cantilevers

Nørlykke, Simon F.; Flyvbjerg, Henrik

2010-01-01

characteristic frequency and the diffusion coefficient. We give analytical results for the weight-dependent bias for the wide class of systems whose dynamics is described by a linear (integro)differential equation with additive noise, white or colored. Examples are optical tweezers with hydrodynamic self...

10. Monitoring Dynamic Protein Expression in Single Living E. Coli. Bacterial Cells by Laser Tweezers Raman Spectroscopy

Chan, J W; Winhold, H; Corzett, M H; Ulloa, J M; Cosman, M; Balhorn, R; Huser, T

2007-01-09

Laser tweezers Raman spectroscopy (LTRS) is a novel, nondestructive, and label-free method that can be used to quantitatively measure changes in cellular activity in single living cells. Here, we demonstrate its use to monitor changes in a population of E. coli cells that occur during overexpression of a protein, the extracellular domain of myelin oligodendrocyte glycoprotein (MOG(1-120)) Raman spectra were acquired of individual E. coli cells suspended in solution and trapped by a single tightly focused laser beam. Overexpression of MOG(1-120) in transformed E. coli Rosetta-Gami (DE3)pLysS cells was induced by addition of isopropyl thiogalactoside (IPTG). Changes in the peak intensities of the Raman spectra from a population of cells were monitored and analyzed over a total duration of three hours. Data was also collected for concentrated purified MOG(1-120) protein in solution, and the spectra compared with that obtained for the MOG(1-120) expressing cells. Raman spectra of individual, living E. coli cells exhibit signatures due to DNA and protein molecular vibrations. Characteristic Raman markers associated with protein vibrations, such as 1257 cm{sup -1}, 1340 cm{sup -1}, 1453 cm{sup -1} and 1660 cm{sup -1}, are shown to increase as a function of time following the addition of IPTG. Comparison of these spectra and the spectra of purified MOG protein indicates that the changes are predominantly due to the induction of MOG protein expression. Protein expression was found to occur mostly within the second hour, with a 470% increase relative to the protein expressed in the first hour. A 230% relative increase between the second and third hour indicates that protein expression begins to level off within the third hour. It is demonstrated that LTRS has sufficient sensitivity for real-time, nondestructive, and quantitative monitoring of biological processes, such as protein expression, in single living cells. Such capabilities, which are not currently available in

11. Impairment of cytoskeleton-dependent vesicle and organelle translocation in green algae: combined use of a microfocused infrared laser as microbeam and optical tweezers.

Holzinger, A; Monajembashi, S; Greulich, K O; Lütz-Meindl, U

2002-11-01

A Nd-YAG laser at 1064 nm is used as optical tweezers to move intracellular objects and a laser microbeam to cause impairment of cytoskeleton tracks and influence intracellular motions in desmidiaceaen green algae. Naturally occurring migrations of large nuclei are inhibited in Micrasterias denticulata and Pleurenterium tumidum when the responsible microtubules are targeted with a laser microbeam generating 180 mW power in the focal plane. Impairment of the microtubule tracks appears to be irreversible, as the nucleus cannot pass the former irradiated area in Pleurenterium or remains abnormally dislocated in Micrasterias. The actin filament-dependent movement of secretory vesicles and smaller particles can be manipulated by the same IR-laser at 90 mW when functioning as optical tweezers. In Closterium lunula particles are displaced from their cytoplasmic tracks for up to 10 micro m but return to their tracks immediately after removing the light pressure gained by the optical tweezers. The cytoplasmic tracks consist of actin filament cables running parallel to the longitudinal axis of Closterium cells as depicted by Alexa phalloidin staining and confocal laser scanning microscopy. Dynamics and extensibility of the cytoplasmic strands connecting particles to the tracks are also demonstrated in the area of large vacuoles which are surrounded by actin filament bundles. In Micrasterias trapping of secretory vesicles by the optical tweezers causes irreversible malformations of the cell shape. The vesicle accumulation itself dissipates within 30 s after removing the optical tweezers, also indicating reversibility of the effects induced, in the case of actin filament-mediated processes. PMID:12423258

12. tweezercalib 2.0: Faster version of MatLab package for precise calibration of optical tweezers

Hansen, Poul Martin; Tolić-Nørrelykke, Iva Marija; Flyvbjerg, Henrik; Berg-Sørensen, Kirstine

2006-03-01

We present a vectorized version of the MatLab (MathWorks Inc.) package tweezercalib for calibration of optical tweezers with precision. The calibration is based on the power spectrum of the Brownian motion of a dielectric bead trapped in the tweezers. Precision is achieved by accounting for a number of factors that affect this power spectrum, as described in vs. 1 of the package [I.M. Tolić-Nørrelykke, K. Berg-Sørensen, H. Flyvbjerg, Matlab program for precision calibration of optical tweezers, Comput. Phys. Comm. 159 (2004) 225-240]. The graphical user interface allows the user to include or leave out each of these factors. Several "health tests" are applied to the experimental data during calibration, and test results are displayed graphically. Thus, the user can easily see whether the data comply with the theory used for their interpretation. Final calibration results are given with statistical errors and covariance matrix. New version program summaryTitle of program: tweezercalib Catalogue identifier: ADTV_v2_0 Program summary URL:http://cpc.cs.qub.ac.uk/summaries/ADTV_v2_0 Program obtainable from: CPC Program Library, Queen's University of Belfast, N. Ireland Reference in CPC to previous version: I.M. Tolić-Nørrelykke, K. Berg-Sørensen, H. Flyvbjerg, Comput. Phys. Comm. 159 (2004) 225 Catalogue identifier of previous version: ADTV Does the new version supersede the original program: Yes Computer for which the program is designed and others on which it has been tested: General computer running MatLab (Mathworks Inc.) Operating systems under with the program has been tested: Windows2000, Windows-XP, Linux Programming language used: MatLab (Mathworks Inc.), standard license Memory required to execute with typical data: Of order four times the size of the data file High speed storage required: none No. of lines in distributed program, including test data, etc.: 135 989 No. of bytes in distributed program, including test data, etc.: 1 527 611 Distribution

13. Molecular cleft or tweezer compounds derived from trioxabicyclo[3.3.1]nonadiene diisocyanate and diacid dichloride

Gert Kollenz

2015-01-01

Full Text Available The structures of two derivatives of the bisdioxine diisocyanate 1, the bisurea 4 and the biscarbamate 5, are established by X-ray crystallography and DFT calculations. These compounds possess endo,endo structures, in the case of the bisurea 4 with two nearly parallel pendant chains. The X-ray structures are reproduced very well by DFT calculations. Similar endo,endo conformations are calculated for the bisamide crown ether derivatives 7, where two proximate and nearly parallel crown ether units endow the molecules with a claw-like molecular cleft or tweezer structure as evidenced by an enhanced ability to extract some alkali, alkaline earth and rare earth metal ions.

14. Parallel optical tweezers with combining a diffractive optical element and a spatial light modulator for photonic DNA memory

Zheng, MingJie; Tate, Naoya; Ogura, Yusuke; Tanida, Jun

2007-09-01

To overcome the restriction of the density of optical memory systems due to diffraction limit, we have been studying photonic DNA memory, which utilizes photonic technologies and the DNA computing methodology. Our scheme is on the basis of local DNA reaction using laser irradiation and transportation of DNA using parallel optical tweezers with fabricating DNA clusters by attaching DNA onto beads. This paper reports on a new dynamic optical tweezers system, which combines a spatial light modulator (SLM) and a diffractive optical element (DOE) for manipulating DNA clusters. With this combination, real-time programmable manipulation of DNA clusters is achievable in a large spatial range. We also can choose simple patterns for the SLM, and decrease computation cost. In this experiment, a laser beam (633nm wavelength) illuminates a SLM (Hamamatsu Photonics K. K.; PPM8267), which is imaged on an 80-lp/mm transmission-type grating, then the beam is focused with a water immersible objective lens (x 100, NA 1). Simple blazed-phase patterns have different grating constants that are perpendicular to that of the grating are displayed on the SLM. We succeeded in lifting up three 6-micron-diameter polystyrene beads on a glass slide with light spots duplicated by the grating, then transporting the beads in three dimensions simultaneously with changing the grating constants on the SLM. We demonstrated that a same manipulation was implemented at different positions by duplicating a pattern that was generated when only using the SLM. This is usable in implementing a same operation for different data at multiple positions with a single instruction. The promising applications of the method include a nano-scale image memory with encryption.

15. Laser microbeams for DNA damage induction, optical tweezers for the search on blood pressure relaxing drugs: contributions to ageing research

Grigaravicius, P.; Monajembashi, S.; Hoffmann, M.; Altenberg, B.; Greulich, K. O.

2009-08-01

One essential cause of human ageing is the accumulation of DNA damages during lifetime. Experimental studies require quantitative induction of damages and techniques to visualize the subsequent DNA repair. A new technique, the "immuno fluorescent comet assay", is used to directly visualize DNA damages in the microscope. Using DNA repair proteins fluorescently labeled with green fluorescent protein, it could be shown that the repair of the most dangerous DNA double strand breaks starts with the inaccurate "non homologous end joining" pathway and only after 1 - 1 Â½ minutes may switch to the more accurate "homologous recombination repair". One might suggest investigating whether centenarians use "homologous recombination repair" differently from those ageing at earlier years and speculate whether it is possible, for example by nutrition, to shift DNA repair to a better use of the error free pathway and thus promote healthy ageing. As a complementary technique optical tweezers, and particularly its variant "erythrocyte mediated force application", is used to simulate the effects of blood pressure on HUVEC cells representing the inner lining of human blood vessels. Stimulating one cell induces in the whole neighbourhood waves of calcium and nitric oxide, known to relax blood vessels. NIFEDIPINE and AMLODIPINE, both used as drugs in the therapy of high blood pressure, primarily a disease of the elderly, prolong the availability of nitric oxide. This partially explains their mode of action. In contrast, VERAPAMILE, also a blood pressure reducing drug, does not show this effect, indicating that obviously an alternative mechanism must be responsible for vessel relaxation.

16. Determining the binding mode and binding affinity constant of tyrosine kinase inhibitor PD153035 to DNA using optical tweezers

Research highlights: → PD153035 is a DNA intercalator and intercalation occurs only under very low salt concentration. → The minimum distance between adjacent bound PD153035 ∼ 11 bp. → Binding affinity constant for PD153035 is 1.18(±0.09) x 104 (1/M). → The change of binding free energy of PD153035-DNA interaction is -5.49 kcal mol-1 at 23 ± 0.5 oC. -- Abstract: Accurately predicting binding affinity constant (KA) is highly required to determine the binding energetics of the driving forces in drug-DNA interactions. Recently, PD153035, brominated anilinoquinazoline, has been reported to be not only a highly selective inhibitor of epidermal growth factor receptor but also a DNA intercalator. Here, we use a dual-trap optical tweezers to determining KA for PD153035, where KA is determined from the changes in B-form contour length (L) of PD153035-DNA complex. Here, L is fitted using a modified wormlike chain model. We found that a noticeable increment in L in 1 mM sodium cacodylate was exhibited. Furthermore, our results showed that KA = 1.18(±0.09) x 104 (1/M) at 23 ± 0.5 oC and the minimum distance between adjacent bound PD153035 ∼ 11 bp. We anticipate that by using this approach we can determine the complete thermodynamic profiles due to the presence of DNA intercalators.

17. Axial and transverse acoustic radiation forces on a fluid sphere placed arbitrarily in Bessel beam standing wave tweezers

Mitri, F. G.

2014-03-01

The axial and transverse radiation forces on a fluid sphere placed arbitrarily in the acoustical field of Bessel beams of standing waves are evaluated. The three-dimensional components of the time-averaged force are expressed in terms of the beam-shape coefficients of the incident field and the scattering coefficients of the fluid sphere using a partial-wave expansion (PWE) method. Examples are chosen for which the standing wave field is composed of either a zero-order (non-vortex) Bessel beam, or a first-order Bessel vortex beam. It is shown here, that both transverse and axial forces can push or pull the fluid sphere to an equilibrium position depending on the chosen size parameter ka (where k is the wave-number and a the sphere's radius). The corresponding results are of particular importance in biophysical applications for the design of lab-on-chip devices operating with Bessel beams standing wave tweezers. Moreover, potential investigations in acoustic levitation and related applications in particle rotation in a vortex beam may benefit from the results of this study.

18. Probing the effect of elevated cholesterol on the mechanical properties of membrane-cytoskeleton by optical tweezers

Rajkumar, Arun S.; Muley, Ajit; Chatterjee, Suvro; Jaffar Ali, B. M.

2010-08-01

The composition of the cell membrane and the surrounding physiological factors determine the nature and dynamics of membrane-cytoskeleton coupling. Mechanical strength of a cell is mainly derived from such coupling. In this article, we investigate the effect of extra cellular cholesterol on the membrane-cytoskelaton connectivity of single cell endothelium and consequent remodeling of its mechanical properties. Using optical tweezers as a force probe, we have measured membrane stiffness (km), membrane microviscosity (ηeff ) and the two-dimensional shear modulus (G'(f)) as a function of extracellular cholesterol in the range of 0.1mM to 6mM. We find that membrane stiffness and shear modulus are dependent on cholesterol-induced membrane-cytoskeletal organization. Further, by disrupting the membranecytoskeletal connectivity with Cytochalasin D, an actin delpolymerizing molecule, we recover pure membrane behaviour devoid of any cytoskeleton attachment. However, behaviour of ηeff was found to be unaffected by disruption of membrane-cytoskeleton organization. We infer that cholesterol is playing a distinct role in modulating membrane organization and membrane-cytoskeleton connectivity independently. We further discuss implications of our approach in characterizing cellular mechanics.

19. Measurements and modeling of water transport and osmoregulation in a single kidney cell using optical tweezers and videomicroscopy

Lúcio, A. D.; Santos, R. A.; Mesquita, O. N.

2003-10-01

With an optical tweezer installed in our optical microscope we grab a single Madin Darby Canine kidney cell and keep it suspended in the medium without touching the glass substrate or other cells. Since the optically trapped cell remains with a closely round shape, we can directly measure its volume by using videomicroscopy with digital image analysis. We submit this cell to a hyperosmotic shock (up-shock) and video record the process: the cell initially shrinks due to osmotic efflux of water and after a while, due to regulatory volume increase (RVI), an osmoregulation response, it inflates again (water influx) until it reaches a new volume (the regulatory volume VR). In addition to considering standard osmotic water transport, we model RVI using a simple phenomenological model. We obtain an expression for cell volume variation as a function of time that fits very well with our experimental data, where two characteristic times appear naturally: one related to water transport and the other related to RVI. From the fit we obtain water permeability, osmolyte influx rate for RVI, and regulatory volume. With the addition of the hormone vasopressin, water permeability increases while the regulatory volume decreases until inhibition of RVI. In summary, we present a technique to measure directly volume changes of a single isolated kidney cell under osmotic shock and a phenomenological analysis of water transport that takes into account osmoregulation.

20. FACS-sorted particles reduce the data variance in optical tweezers-assisted dynamic force spectroscopy measurements

By combining optical tweezers-assisted dynamic force spectroscopy experiments with fluorescence activated cell sorting (FACS), we demonstrate a new approach to reducing the data variance in measuring receptor–ligand interactions on a single molecule level by ensuring similar coating densities. Therefore, the carboxyfluorescein-labelled monophosphorylated peptide tau226–240[pThr231] is anchored on melamine resin beads and these beads are sorted by FACS to achieve a homogeneous surface coverage. To quantify the impact of the fluorescence dye on the bond parameters between the phosphorylated peptide and the corresponding phosphorylation specific anti-human tau monoclonal antibody HPT-104, we perform dynamic force spectroscopy and compare the results to data using unsorted beads covered with the non-fluorescence peptide analogue. Finally, we demonstrate that the data variance of the relative binding frequency is significantly decreased by a factor of 3.4 using pre-sorted colloids with a homogeneous ligand coating compared to using unsorted colloids. (paper)

1. Fast characterisation of cell-derived extracellular vesicles by nanoparticles tracking analysis, cryo-electron microscopy, and Raman tweezers microspectroscopy

Irène Tatischeff

2012-11-01

Full Text Available The joint use of 3 complementary techniques, namely, nanoparticle tracking analysis (NTA, cryo-electron microscopy (Cryo-EM and Raman tweezers microspectroscopy (RTM, is proposed for a rapid characterisation of extracellular vesicles (EVs of various origins. NTA is valuable for studying the size distribution and concentration, Cryo-EM is outstanding for the morphological characterisation, including observation of vesicle heterogeneity, while RTM provides the global chemical composition without using any exogenous label. The capabilities of this approach are evaluated on the example of cell-derived vesicles of Dictyostelium discoideum, a convenient general model for eukaryotic EVs. At least 2 separate species differing in chemical composition (relative amounts of DNA, lipids and proteins, presence of carotenoids were found for each of the 2 physiological states of this non-pathogenic microorganism, that is, cell growth and starvation-induced aggregation. These findings demonstrate the specific potency of RTM. In addition, the first Raman spectra of human urinary exosomes are reported, presumably constituting the primary step towards Raman characterisation of EVs for the purpose of human diseases diagnoses.

2. Disrupting self-assembly and toxicity of amyloidogenic protein oligomers by "molecular tweezers" - from the test tube to animal models.

Attar, Aida; Bitan, Gal

2014-01-01

Despite decades of research, therapy for diseases caused by abnormal protein folding and aggregation (amyloidoses) is limited to treatment of symptoms and provides only temporary and moderate relief to sufferers. The failure in developing successful disease-modifying drugs for amyloidoses stems from the nature of the targets for such drugs - primarily oligomers of amyloidogenic proteins, which are distinct from traditional targets, such as enzymes or receptors. The oligomers are metastable, do not have well-defined structures, and exist in dynamically changing mixtures. Therefore, inhibiting the formation and toxicity of these oligomers likely will require out-of-the-box thinking and novel strategies. We review here the development of a strategy based on targeting the combination of hydrophobic and electrostatic interactions that are key to the assembly and toxicity of amyloidogenic proteins using lysine (K)-specific "molecular tweezers" (MTs). Our discussion includes a survey of the literature demonstrating the important role of K residues in the assembly and toxicity of amyloidogenic proteins and the development of a lead MT derivative called CLR01, from an inhibitor of protein aggregation in vitro to a drug candidate showing effective amelioration of disease symptoms in animal models of Alzheimer's and Parkinson's diseases. PMID:23859557

3. Developing a New Biophysical Tool to Combine Magneto-Optical Tweezers with Super-Resolution Fluorescence Microscopy

Zhaokun Zhou

2015-06-01

Full Text Available We present a novel experimental setup in which magnetic and optical tweezers are combined for torque and force transduction onto single filamentous molecules in a transverse configuration to allow simultaneous mechanical measurement and manipulation. Previously we have developed a super-resolution imaging module which, in conjunction with advanced imaging techniques such as Blinking assisted Localisation Microscopy (BaLM, achieves localisation precision of single fluorescent dye molecules bound to DNA of ~30 nm along the contour of the molecule; our work here describes developments in producing a system which combines tweezing and super-resolution fluorescence imaging. The instrument also features an acousto-optic deflector that temporally divides the laser beam to form multiple traps for high throughput statistics collection. Our motivation for developing the new tool is to enable direct observation of detailed molecular topological transformation and protein binding event localisation in a stretching/twisting mechanical assay that previously could hitherto only be deduced indirectly from the end-to-end length variation of DNA. Our approach is simple and robust enough for reproduction in the lab without the requirement of precise hardware engineering, yet is capable of unveiling the elastic and dynamic properties of filamentous molecules that have been hidden using traditional tools.

4. Hybrid optical-electrochemical electronic nose system based on Zn-porphyrin and multi-walled carbon nanotube composite.

Kladsomboon, Sumana; Lutz, Mario; Pogfay, Tawee; Puntheeranurak, Theeraporn; Kerdcharoen, Teerakiat

2012-07-01

In this work, we have enhanced the capability of an e-nose system based on combined optical and electrochemical transduction within a single gas sensor array. The optical part of this e-nose is based on detection of the absorption changes of light emitted from eight light emitting diodes (LEDs) as measured by a CMOS photo-detector. The electrochemical part works by measuring the change in electrical resistivity of the sensing materials upon contact with the sample vapor. Zinc-5,10,15,20-tetra-phenyl-21H,23H-porphyrin (ZnTPP) and multi-walled carbon nanotube (MWCNT) composite was used as the sensing materials based on its good optoelectronic properties. This sensing layer was characterized by UV-Vis spectroscopy and atomic force microscope and tested with various VOC vapors. Density functional theory (DFT) calculations were performed to investigate the electronic properties and interaction energies between ZnTPP and analyte molecules. It can be clearly seen that this hybrid optical-electrochemical electronic nose system can classify the vapor of different volatile organic compounds. PMID:22966552

5. The study of adhesive forces between the type-3 fimbriae of Klebsiella pneumoniae and collagen-coated surfaces by using optical tweezers

Chan, Chiahan; Fan, Chia-chieh; Huang, Ying-Jung; Peng, Hwei-Ling; Long, Hsu

2004-10-01

Adherence to host cells by a bacterial pathogen is a critical step for establishment of infection. It will contribute greatly to the understanding of bacterial pathogenesis by studying the biological force between a single pair of pathogen and host cell. In our experiment, we use a calibrated optical tweezers system to detach a single Klebsiella pneumoniae, the pathogen, from collagen, the host. By gradually increasing the laser power of the optical tweezers until the Klebsiella pneumoniae is detached from the collagen, we obtain the magnitude of the adhesive force between them. This happens when the adhesive force is barely equal to the trapping force provided by the optical tweezers at that specific laser power. This study is important because Klebsiella pneumoniae is an opportunistic pathogen which causes suppurative lesions, urinary and respiratory tract infections. It has been proved that type 3 fimbrial adhesin (mrkD) is strongly associated with the adherence of Klebsiella pneumoniae. Besides, four polymorphic mrkD alleles: namely, mrkDv1, v2, v3, and v4, are typed by using RFLP. In order to investigate the relationship between the structure and the function for each of these variants, DNA fragments encoding the major fimbrial proteins mrkA, mrkB, mrkC are expressed together with any of the four mrkD adhesins in E. coli JM109. Our study shows that the E. coli strain carrying the mrkDv3 fimbriae has the strongest binding activity. This suggests that mrkDv3 is a key factor that enhances the adherence of Klebsiella Pneumoniae to human body.

6. Leishmania amazonensis chemotaxis under glucose gradient studied by the strength and directionality of forces measured with optical tweezers

de Ysasa Pozzo, Liliana; Fontes, Adriana; de Thomaz, André A.; Barbosa, Luiz Carlos; Ayres, Diana Copi; Giorgio, Selma; Cesar, Carlos Lenz

2007-02-01

Chemotaxis is the mechanism microorganisms use to sense the environment surrounding them and to direct their movement towards attractive, or away from the repellent, chemicals. The biochemical sensing is almost the only way for communication between unicellular organisms. Prokaryote and Eukaryote chemotaxis has been mechanically studied mainly by observing the directionality and timing of the microorganisms movements subjected to a chemical gradient, but not through the directionality and strength of the forces it generates. To observe the vector force of microorganisms under a chemical gradient we developed a system composed of two large chambers connected by a tiny duct capable to keep the chemical gradient constant for more than ten hours. We also used the displacements of a microsphere trapped in an Optical Tweezers as the force transducer to measure the direction and the strength of the propulsion forces of flagellum of the microorganism under several gradient conditions. A 9μm diameter microsphere particle was trapped with a Nd:YAG laser and its movement was measured through the light scattered focused on a quadrant detector. We observed the behavior of the protozoa Leishmania amazonensis (eukaryote) under several glucose gradients. This protozoa senses the gradient around it by swimming in circles for three to five times following by tumbling, and not by the typical straight swimming/tumbling of bacteria. Our results also suggest that force direction and strength are also used to control its movement, not only the timing of swimming/tumbling, because we observed a higher force strength clearly directed towards the glucose gradient.

7. The recognition mechanism of the 1, 3-di(1-phenyl-1H-tetrazole-5-thiol)-acetylphenylhydrazone tweezer receptor for the halogen anions

The recognition mechanism of the 1, 3-di(1-phenyl-1H-tetrazole-5-thiol)-acetylphenylhydrazone tweezer receptor (DAPHZ) for the halogen anions was discussed at the density function Becke, three-parameter, Lee-Yang-Parr (B3LYP) and Hatree-Fock (HF) methods. The results showed that recognition mechanism was performed by using red-shift hydrogen bond between N-H bonds of the DAPHZ and the halogen anions X- (X=F-, Cl- and Br-). The calculated interaction energies (ΔECP) with basis set super-position error (BSSE) correction of the three complexes are -327.5, -163.5 and -148.3 kJmol-1 (HF/3-21G), respectively. Natural bond orbital theory (NBO) analysis, Atoms in molecules (AIM) theory and the chemical shift calculation of the related atoms has been used to investigate the electronic structure and properties of the red-shift hydrogen bond between N-H bonds of the DAPHZ and the halogen anions X-, and the results revealed that the relative recognition ability of the DAPHZ tweezer receptor for the halogen anions are in the order of DAPHZ…F- >>DAPHZ…Cl-∼DAPHZ…Br-. (authors)

8. Fluorescence Detection of H5N1 Virus Gene Sequences Based on Optical Tweezers with Two-Photon Excitation Using a Single Near Infrared Nanosecond Pulse Laser.

Li, Cheng-Yu; Cao, Di; Kang, Ya-Feng; Lin, Yi; Cui, Ran; Pang, Dai-Wen; Tang, Hong-Wu

2016-04-19

We present an analytical platform by combining near-infrared optical tweezers with two-photon excitation for fluorescence detection of H5N1 virus gene sequences. A heterogeneous enrichment strategy, which involved polystyrene (PS) microsphere and quantum dots (QDs), was adopted. The final hybrid-conjugate microspheres were prepared by a facile one-step hybridization procedure by using PS microspheres capturing target DNA and QDs tagging, respectively. Quantitative detection was achieved by the optical tweezers setup with a low-cost 1064 nm nanosecond pulse laser for both optical trapping and two-photon excitation for the same hybrid-conjugate microsphere. The detection limits for both neuraminidase (NA) gene sequences and hemagglutinin (HA) gene sequences are 16-19 pM with good selectivity for one-base mismatch, which is approximately 1 order of magnitude lower than the most existing fluorescence-based analysis method. Besides, because of the fact that only signal from the trapped particle is detected upon two-photon excitation, this approach showed extremely low background in fluorescence detection and was successfully applied to directly detect target DNA in human whole serum without any separation steps and the corresponding results are very close to that in buffer solution, indicating the strong anti-interference ability of this method. Therefore, it can be expected to be an emerging alternative for straightforward detecting target species in complex samples with a simple procedure and high-throughput. PMID:27023254

9. To Study the Effect of Paclitaxel on the Cytoplasmic Viscosity of Murine Macrophage Immune Cell RAW 264.7 Using Self-Developed Optical Tweezers System

Chen, Ying-chun; Wu, Chien-ming

2012-12-01

In recent years, optical tweezers have become one of the tools to measure the mechanical properties of living cells. In this study, we first constructed an optical tweezers to investigate the cytoplasmic viscosity of immune cells. In addition to measuring viscosity of cells in a normal condition, we also treated cells with anti-cancer drug, Paclitaxel, and in order to study its effect on the cytoplasmic viscosity. The results showed that the viscosity decreased dramatically during the first 3 h. After 3 h, the change started to slow down and it remained nearly flat by the end of the experiment. In addition, we used the confocal laser scanning microscope to observe the cytoskeleton of the cell after drug treatment for 3 and 5 h, respectively, and found that actin filaments were disrupted and that the nucleus had disintegrated in some drug-treated cells, similar to the process of apoptosis. This study presents a new way for measuring the changes in cytoplasmic viscosity, and to determine if a cell is going into apoptosis as a result of a drug treatment.

10. Imaging and spectroscopic analysis of single microdroplets containing p-cresol using the near-infrared laser tweezers/Raman microprobe system

Ajito, Katsuhiro; Morita, Masao

1999-06-01

The near-infrared (NIR) laser tweezers/Raman microprobe system features two charge-coupled device (CCD) cameras with holographic notch filters (HNFs) for the imaging and spectroscopic analysis of molecules in a single microdroplet (MD). One CCD camera and a HNF are used to record an image of the laser microprobe in a trapped MD. The other CCD camera and two HNFs are used with a polychromator to obtain a Raman spectrum of molecules in the MD. A dielectric multilayer coated beam splitter divides the scattered NIR light into two optical paths for the cameras. The system provides sufficient sensitivity to obtain a Raman spectrum of p-cresol contained in a single picoliter toluene MD and sufficient spatial resolution to record an image of the laser microprobe in a trapped MD simultaneously. Furthermore, a difference in the solubility for the p-cresol in bulk solvent and in the MD solvent was clearly observed using this system.
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