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A side-entry excitation arrangement is provided with a multi-channel analyte-separation device. In various embodiments, a plurality of channels are disposed in an array, with a laser disposed to direct an excitation beam of light along a beam path that crosses the longitudinal axes of the channels, so as to simultaneously irradiate a region of each of the channels. Devices of the invention can be useful, for example, in the separation and analysis of bio-molecules, such as DNA, RNA, etc.
1. A device for the analysis of one or more samples, comprising:
a substrate;
a plurality of adjacently arranged channels formed in said substrate, with each channel having an inlet end and an outlet end; said channels being disposed in spaced relation relative to one another, with each adjacent pair of channels being separated by a respective portion of said substrate that includes at least a region that is transparent; and
an excitation-beam source adapted to direct an excitation beam of light along a beam path that intersects each of said channels at a region between said inlet and outlet ends and further intersects the transparent region of the substrate separating adjacent pairs of channels.
2. The device of
3. The device of
4. The device of
5. The device of
6. The device of
7. The device of
8. The device of
9. A device for the analysis of one or more samples, comprising:
a substrate, including one or more transparent regions;
a plurality of adjacently arranged, elongate separation channels formed in said substrate, with each adjacent pair of said separation channels being separated, at least in part, by at least a portion of said one or more transparent regions; and
an excitation-beam source disposed to direct an excitation beam of light along a beam path intersecting each of said separation channels and said at least portions of said one or more transparent regions.
10. The device of
11. The device of
12. A device for the analysis of one or more samples, including:
a substrate including a plurality of adjacently arranged, non-intersecting separation channels, each channel comprising an inlet end and an outlet end;
a wall structure separating each adjacent pair of channels, each wall structure including at least a portion that is transparent to a limited wavelength range of light;
an excitation-beam path extending across the substrate, intersecting (i) each channel at a region between its inlet and outlet ends and (ii) each transparent portion of said wall structure.
13. The device of
an excitation beam source configured to direct an excitation beam of light along said excitation-beam path, said excitation beam of light being within said limited wavelength range.
14. The device of
an emission detection system optically coupled to a region within each channel along said excitation-beam path.
The present invention relates to electrophoretic separation systems for the analysis of bio-molecules, such as nucleic acids. More particularly, this invention relates to a multi-channel analyte-separation device employing side-entry illumination.
Backhouse et al., DNA sequencing in a monolithic microchannel device, Electrophoresis 2000, 21, 150-156.
Dolnik et al., Capillary electrophoresis on microchip, Electrophoresis 2000, 21, 41-54.
Grossman and Colburn, Capillary Electrophoresis Theory and Practice, Chapter 1, Academic Press (1992).
Kambara et al., U.S. Pat. No. 5,192,142 (1993).
Madabhushi et al., U.S. Pat. No. 5,552,028 (1996).
Sambrook et al., eds., Molecular Cloning: A Laboratory Manual, Second Edition, Chapter 5, Cold Spring Harbor Laboratory Press (1989).
Woolley et al., Ultra-high-speed DNA fragment separations using microfabricated capillary array electrophoresis chips, Proc. Natl. Acad. Sci., vol. 91, pp. 11348-11352, November 1994, Biophysics.
Yeung et al., U.S. Pat. Nos. 5,741,411 and 5,582,705.
Devices for carrying out separations of analytes, such as biomolecules (e.g., proteins, DNA, RNA, etc.), have gained widespread use in recent years.
In electrophoretic separations, it is often desirable to illuminate a plurality of migrating analytes, tagged with excitable reporters (e.g., fluorescent dyes), to stimulate detectable emission indicative of the nature (e.g., identity or composition) of the tagged analytes.
Various aspects of the present invention provide a multi-channel analyte-separation device (channel device) comprising a substrate defining an array of channels. According to various embodiments, adjacent channels of the device are separated by wall structure, which includes at least a portion that is substantially transparent. The transparent portions are disposed along a path or line crossing (e.g., co-planar and normal to) the longitudinal axes of the channels. An excitation-beam source (e.g., a laser) is adapted to direct an excitation beam of light along the path, such that the beam can simultaneously pass through each of the transparent portions and each of the channels. Plural samples migrating along the various channels, e.g., by electrophoresis, can thus be simultaneously irradiated and detected.
Various embodiments are particularly adapted to bio-molecule (e.g., DNA, RNA, PNA, etc.) sequence or other analysis methods, in which each of a plurality of different fragment types is labeled with a spectrally distinctive fluorescent dye. According to certain embodiments, a side-entry laser arrangement at a detection zone of a multi-channel electrophoresis device excites the dyes, while in the channels, to emit light. In various embodiments, emitted light from samples in the channels passes through a laser light filter, through a collection lens, through a transmission dispersion element, which spectrally separates the light, and through a focusing lens. The focused light can be incident on a detector array (e.g., CCD) capable of detecting the simultaneously spatially focused and spectrally diverged light from the detection regions of all the channels. Electronic signals from the detector array can provide information about the character or sequence of the DNA sample.
The structure and manner of operation of the invention may further be understood by reference to the following description taken in conjunction with the accompanying drawings, in which identical reference numerals identify identical or similar elements, and in which:
FIG. 1 is a perspective view from above of an electrophoresis system, showing a multi-channel analyte-separation device including a plurality of separation channels, a detection zone, an excitation beam source, an optical detection system, and a programmed computer control/analysis system, according to various embodiments;
FIGS. 2A, 2B, and 2C are partial, cross-sectional views of multi-channel analyte-separation devices, according to various embodiments;
FIGS. 3A, 3B, and 3C are cross-sectional views of substrates with channels formed therein having various geometries, according to certain embodiments; and
FIG. 4 is a perspective view from above of an electrophoresis system, including a multi-channel analyte-separation device including a plurality of separation channels, an excitation beam source, and optics directing an excitation beam for entry into the channel device from each lateral side thereof, according to various embodiments.
Reference will now be made to various embodiments of the invention, examples of which are illustrated in the accompanying drawings. While the invention will be described in conjunction with various preferred embodiments, it will be understood that they are not intended to limit the invention. On the contrary, the invention is intended to cover alternatives, modifications, and equivalents, which may be included within the invention as defined by the appended claims.
Unless stated otherwise, the following terms and phrases as used herein are intended to have the following meanings:
The term "channel" as used herein refers to an elongate, narrow passage or other structure (e.g., grooves, etc.) formed in a substrate and capable of supporting a volume of separation medium and/or buffer solution; e.g., such as is used in carrying out electrophoresis. The geometry of a channel may vary widely. For example, a channel can have a circular, oval, semi-circular, semi-oval, triangular, rectangular, square, or other cross-section, or a combination thereof. Channels can be fabricated by a wide range of technologies, including microfabrication techniques. As used herein, the term "channel" is not intended to encompass a capillary tube.
The terms "capillary" and "capillary tube" as used herein, refer to an elongated tubular or cylindrical structure defining an inner lumen. For example, a capillary can be an elongated capillary or micro-capillary tube made, for example, from fused silica, quartz, silicate-based glass, such as borosilicate glass, phosphate glass, alumina-containing glass, and the like, or other silica-like material(s). As used herein, "capillary" does not encompass a channel in a substrate such as a plate, slide, chip, wafer, or the like.
The term "channel device" refers to a substrate, such as a plate, slide, chip, wafer, or similar structure, including one or more channels (e.g., grooves); and particularly those adapted at least in part for carrying out electrophoresis. Channel devices can take the form, for example, of microfabricated devices (e.g., a grooved, etched, or fluted plate, slide, chip, wafer, or other substrate).
As used herein, the term "sample zone" or "analyte zone" refers to a collection of molecules comprising a subset of sample or analyte components having similar electrophoretic migration velocities such that the molecules of a sample zone or analyte zone migrate as a defined zone. In the limit, such a zone is made up of molecules having identical electrophoretic migration velocities. Sample zones and analyte zones are often referred to as "bands."
As used herein, the term "separation medium" or "separation matrix" refers to a medium in which an electrophoretic separation of sample components can take place. Separation media typically comprise several components, at least one of which is a charge-carrying component, or electrolyte. The charge-carrying component is usually part of a buffer system for maintaining the separation medium at a defined pH. Media for separating polynucleotides, proteins, or other biomolecules having different sizes but identical charge-frictional drag ratios in free solution, further include a sieving component. Such sieving component is typically composed of a cross-linked polymer gel, e.g., cross-linked polyacrylamide or agarose (Sambrook), or a polymer solution, e.g., a solution of polyacrylamide, hydroxyethyl cellulose, and the like (Grossman; Madabhushi).
Various aspects of the present invention provide channel devices useful, for example, in electrophoretic separations of bio-molecules. According to various embodiments, the channel devices employ side-entry excitation geometry. Channel devices herein are to be contrasted with capillary arrangements that employ multiple capillaries (i.e., elongated tubular structures). Rather, the channel devices herein are comprised of a substrate, such as a plate, slide, chip, wafer, or similar structure, including one or more channels (e.g., grooves). The substrate can be a monolithic structure or a multi-layer structure, according to one or more embodiments. In various embodiments, channel devices take the form, for example, of microfabricated devices (e.g., grooved, etched, or fluted plate, slide, chip, wafer, or other substrate). It has previously been suggested by others that channel-device technology was not developed enough to employ side-entry illumination (see Yeung et al., U.S. Pat. No. 5,741,411, col. 8, lines 14-24, and U.S. Pat. No. 5,582,705, col. 8, lines 9-19). To the contrary, the present invention prefers the use of channel devices over multiple capillary (tube) arrangements.
In various embodiments, separation channels are formed on a glass or plastic substrate, such as a plate, slide, wafer, chip, or the like, by microfabrication techniques known in the art, e.g., photolithographical and/or wet-chemical etching procedures, laser ablation, electroforming, microcontact printing, microstamping, micromolding, microcasting, micromachining, engraving, and/or embossing techniques, to name a few. For example, Backhouse et al., Dolnik et al., and Woolley et al (each of which is incorporated herein by reference) discuss certain fabrication techniques that the skilled artisan can employ in making the devices of the present invention. In one embodiment, the separation channels are formed in a generally planar substrate comprised at least in part, for example, of an electrically insulating material, e.g., fused silica, quartz, silicate-based glass, such as borosilicate glass, phosphate glass, alumina-containing glass, and the like, or other silica-like material(s).
Various embodiments of the devices herein are particularly well suited, for example, to fluorescence detection of a fluorescent target species in a sample. According to various embodiments, channels of a channel device are arranged in a coplanar channel array. In various embodiments, the channel array includes at least about 4 (e.g., 8, 12, 16, 24, 48, 96, or more) coplanar, adjacently arranged channels. Sidewall regions of each channel include one or more transparent portions. According to certain embodiments, a transparent portion is transparent to light having a wavelength about equal to a wavelength of a beam of coherent light used to irradiate a target species in a channel. A "transparent portion" or "transparent medium" is one that transmits light with little or no attendant light scattering. For example, a transparent portion can be comprised of an optically clear glass or plastic. According to certain embodiments, the transparent portion is transparent to light having a wavelength of about 200-1500 nm; e.g., about 250-800 nm.
Together, the transparent portions define a transparent path extending through the channel array, e.g., from each channel to the next. In an embodiment, the transparent path comprises a plane extending through the channels, as is the case where the channels are fabricated entirely out of transparent material.
In certain embodiments, the transparent portions of the sidewalls exhibit little or no fluorescence when exposed to a beam of coherent light, so as to reduce or eliminate background fluorescence from the detected fluorescence. For example, the transparent portions can be selected and designed to exhibit substantially no fluorescence when exposed light having a wavelength of 200-1500 nm; e.g., about 250-800 nm. By "substantially no fluorescence" is meant that the level of fluorescence emitted by a transparent portion, if any, is less than observed background fluorescence.
According to various embodiments, detection of a target species can be effected through a transparent portion provided in an upper wall or ceiling region of a channel. Such additional transparent portion can be selected and designed to exhibit substantially no fluorescence when exposed to light having a wavelength about equal to the wavelength of light emitted by a fluorescing target species. In an embodiment, the entire channel device is constructed from a transparent, non-fluorescing material, such as fused silica. Transparent windows may alternatively be formed at or along selected regions of channels.
Instead of, or in addition to, utilizing such transparent portions, one or more sidewalls can include a translucent portion defining a translucent path extending through the array perpendicular to the channels. A translucent medium produces some light scattering when transmitting light. In certain embodiments, transparency is preferred over translucency because of greater light throughput and reduced detection S/N.
As indicated above, side-entry irradiation of target species in multiple channels can be effected through a transparent portion of a sidewall of each channel in a multi-channel array. According to various embodiments, light passes through the transparent portions in the array in a sequential manner. A coherent light source can be positioned to direct a beam of coherent light along the transparent path. A coherent light source produces light waves traveling together in phase. The light can have, for example, a wavelength of about 200-1,500 nm. For example, the coherent light source used can be a laser. An argon ion laser operating simultaneously at one or more visible lines can be used for excitation, although other light sources and wavelengths can be used. Exemplary excitation wavelengths are 488 nm and 514 nm. A pure output laser, i.e., a laser emitting light of a single wavelength, can be a useful light source. Alternatively, the wavelength of the laser can be chosen by an interference filter or a glass prism.
According to various embodiments, the beam of coherent light is focused and collimated through a collimating focusing lens interposed between the coherent light source and the channel array. For example, the collimated excitation beam can have a diameter of less than about 300 micrometers (e.g., less than about 75 or 50 micrometers) while traversing the channels in the array. In an embodiment, including an array comprising about 96 channels, the array width is less than about 1.5 cm, and a lens with a focal length of about 5-30 cm, e.g., about 10 cm, is used to focus and collimate the beam such that the beam diameter remains less than about 75 micrometers while in the channels.
According to various embodiments, the focused line of the laser is altered with a beam expander in order to more effectively irradiate a large number of channels. For example, the laser beam can be expanded perpendicular to the capillary array. Such lengthening or "fanning out" of the laser line can facilitate positioning of the beam so that all channels are adequately irradiated. The beam can optionally be altered or redirected, as with a mirror, filter, lens, or other optical element, prior to contacting the array. For example, mirrors can be used to provide a convenient means for adjusting the direction of the laser beam to become coplanar with the channel array perpendicular to the channels. The use of mirrors, filters, lenses, etc., or any combination thereof, is optional.
A location external to the channel array to which the transparent path may be optically coupled is to be broadly understood as any point, line, or plane external to the array, including a single pixel, linear array of pixels, or planar array (two-dimensional array) of pixels. For example, the location external to the capillary array can comprise a planar surface parallel or angled with respect to the channel array. The location external to the capillary array can include an optical detector, e.g., capable of detecting fluorescence emission from a target species in a sample in a channel. In an embodiment, the optical detector is a two-dimensional image array detector. For example, the optical detector can be a charge-coupled device (CCD) or a charge-injection device (CID).
Referring now to the drawings, FIG. 1 is a perspective view of an embodiment of an electrophoresis device, indicated generally by the reference numeral 12. Device 12 includes a plurality of separation channels, such as elongate channels 14, with each channel having an inlet end and an outlet end, as indicated at 16 and 18 respectively. A first lead wire 22 connects a power source 20 with a first electrode (not visible in FIG. 1) disposed in electrical communication with the inlet ends of the separation channels; and a second lead wire 24 connects source 20 with a second electrode (not visible in FIG. 1) disposed in electrical communication with the outlet ends of the separation channels. In operation, a voltage is applied between the first and second electrodes, and thereby along the channels, such that a sample zone is transported from the inlet ends, to the outlet ends of the channels, and through an on-channel detection zone, denoted as 30, located between the inlet and outlet ends.
In the arrangement of FIG. 1, device 12 is comprised of upper and lower plates, 26 and 28 respectively, with abutted confronting faces. As shown, lower plate includes end portions, 28 a and 28 c, and lateral side portions, 28 b and 28 d. Lower plate 28 is provided with a plurality of non-intersecting elongate grooves, each of roughly semi-circular or semi-oval cross-section, positioned at regular intervals (e.g., at a pitch of about 250 um) and extending along its upper face (e.g., for a length of about 5 cm); which grooves in part define separation channels 14. The lower face of plate 26 is substantially planar, and, when disposed against plate 28 as shown, further defines channels 14. Particularly, in the illustrated arrangement, the grooves of plate 28 define lower (floor) and sidewalls, or boundaries, of each channel 14 and the lower surface of plate 26 provides an upper wall for each channel 14.
Alternatively, both the upper and lower plates can be provided with complimentary sets of grooves that can be aligned with one another so that corresponding upper and lower grooves cooperate to define a plurality of elongate channels.
Instead of providing grooves in a lower plate which are covered by a planar lower surface of an upper plate, such as shown in FIG. 1, the device of the invention can include an upper plate with grooves formed along its lower surface, which can be placed over a planar upper surface of a lower plate (that is, essentially, the reverse of what is shown in FIG. 1). Moreover, although the device of the invention is illustrated as operating with the major planar surfaces of the plates disposed in a substantially horizontal fashion, the device instead could instead be configured to operate with the plates disposed substantially vertically, or tilted at a desired angle.
While the channels depicted in FIG. 1 are parallel to one another, it should be appreciated that other configurations are possible. In one embodiment, the channels converge toward one end of the device (e.g., the distance separating adjacent channels (i.e., the pitch) becomes smaller along a direction towards the outlet ends). Additionally, it should be noted that the central longitudinal axes of the channels can be straight (as shown), curved, or a combination thereof. In the embodiment of FIG. 1, the flow cross-sectional area (i.e., the cross section taken perpendicular to the direction of sample migration) is substantially the same among the various channels. In particular, the channels of FIG. 1 are all of a uniform depth (measured from the lower face of the upper plate bounding the top region of the channel to the lowermost point, or floor, of the channel groove). Such uniformity can be achieved as the ordinary result of common microfabrication methods employed in constructing the device, such as etching. However, the invention additionally contemplates channels of varying depth; which can be made, for example, by use of a two stage etching process with multiple masks.
In practice, a separation medium is injected (e.g., pressure-filled or vacuum aspirated) or otherwise provided in the separation channels of the invention to effect the electrophoretic separation of the components of the sample(s). It should be noted that the present invention contemplates the use of any suitable injection techniques; e.g., without limitation, electrokinetic injections, hydrodynamic injections, cross tee injectors and double tee injectors, etc., as known and described in the art. In one preferred embodiment, the separation medium is a flowable, non-crosslinked polymer solution.
An excitation-beam source, such as 34, is provided for stimulating emission from sample zones located in detection zone 30. In an embodiment, the light source is a laser, e.g., an argon ion laser, a solid-state laser, or the like; however, any suitable beam source can be used. As described in more detail below, in an embodiment of the present invention, an excitation-beam pathway or path extends through detection zone 30, along which an energy beam 48 generated by the beam source can pass. Such pathway is located between the inlet and outlet ends and extends along a plane defined by the channels (i.e., the pathway is co-planar with the plane of the channels). In an embodiment, the beam pathway is perpendicular to the direction of sample migration across the detection chamber, but this is not critical to the invention (i.e., the pathway can approach at an angle with respect to the direction of sample migration). What is important in this embodiment is that a beam passing along the pathway is capable of simultaneously exciting plural sample zones in respective (different) channels.
In various embodiments, such as shown in the arrangement of FIG. 1, the beam enters a lateral side 28 b of lower plate 28, passes through plate 28 (including each of channels 14), and exits at an opposite lateral side 28 d of plate 28. In other embodiments, a laser dump or sink can be incorporated in the plate, e.g., proximate a region of side 28 d, whereat the beam can terminate after passing through the channels.
As previously mentioned, a first electrode (not visible in FIG. 1) is in electrical communication with inlet ends 16 of separation channels 14. During operation of device 12, the first electrode is maintained at a first voltage V1 using power source 20. Electrical communication between the first electrode and the inlet ends of the separation channels can be established, for example, by providing an electrically conductive solution in a reservoir/loading region 35 of device 14 so that both the inlet ends of the channels and the first electrode are in contact with the conductive solution.
With continued reference to FIG. 1, each of outlet ends 18 communicates a respective channel 14 with a second reservoir 37, which reservoir is located proximate the outlet ends.
The second electrode (not visible in FIG. 1) is in electrical communication with outlet ends 18 of separation channels 14. During operation of device 12, the second electrode is maintained at a second voltage V2, also using power supply 20. Preferably, electrical communication between the second electrode and second reservoir 37 is established by providing an electrically conductive solution in second reservoir 37 such that the second electrode and outlet ends 18 are in contact with the conductive solution.
The electrodes used in the device may be formed from any electrically conducting materials. Preferably, the electrodes are made from a chemically inert material, e.g., platinum, gold, stainless steel, or other relatively inert conductive material. In accordance with one embodiment of the present invention, platinum electrodes are fabricated on the top or bottom plate by RF sputtering and photolithography before the top plate is bonded to the etched bottom plate.
The electrically conductive solution used to establish electrical continuity throughout the system can be any fluid capable of transporting an electrical current. For example, the conductive solution can be an ionic solution, e.g., an aqueous solution containing a dissolved salt. In various embodiments, the conductive solution includes a buffer for stabilizing the pH of the solution. According to certain embodiments, the ionic composition of the conductive solution is the same in the separation channels, each of the electrode reservoirs, and the detection chamber.
To facilitate optical detection of sample zones in the detection zone 30, part or all of upper plate 26 covering detection zone 30 can be formed from a material which efficiently transmits light (i.e., an optically clear material), e.g., glass, quartz, clear plastic, and the like. In addition, to facilitate the introduction of an excitation light beam 48 into the detection zone to excite fluorescence of sample zones therein, part or all of the plate 28 along a region between the beam source 34 and the endmost channel 14 closest thereto can be formed from a material which efficiently transmits light. In various embodiments, the light-transmitting material does not significantly scatter light and has little intrinsic fluorescence.
Further in the embodiment of FIG. 1, a detector 38 is provided for detecting sample zones passing through the detection zone 30. The detector can be any type of detector for detecting emission of any type radiation, e.g., radioactivity, fluorescence, phosphorescence, chemi-luminescence, and the like. In an embodiment, detector 38 is capable of detecting fluorescence from a plurality of locations independently and simultaneously, e.g., a CCD camera, an array of photomultiplier tubes, a diode array, and the like. As illustrated in FIG. 1, detector 38 can be connected to a computer 42 to store, analyze, and display data collected by the detector and/or to control the operation of the detector and other aspects of the device, as desired. For example, computer 42 can be programmed to control power source 20 and/or beam source 34.
It should be appreciated that, in regions of the device where it is not required or desired that radiative emission should be able to pass through, non-optically clear materials may be used, e.g., polymeric materials such as Teflon, silicone, and the like. Of course, the detection zone, as previously mentioned, preferably permits light to pass from each channel to the next channel, and to the detector.
The electrophoresis apparatus of the present invention can also include one or more additional elements typically used to conduct a capillary electrophoresis process, e.g., a temperature control device for controlling the temperature of the separation channels. Details of these and other common features of an operable capillary electrophoresis device can be found in any number of available publications, e.g., Capillary Electrophoresis Theory and Practice, Grossman and Colburn, eds., Academic Press (1992).
Various embodiments provide for reduced crosstalk, while facilitating excitation and detection. Certain embodiments, for example, utilize bandpass filters that transmit light only within a defined spectral band. For example, an excitation filter can be employed (e.g., coated upon or fixed to regions of the lower plate) that allows only light which excites a reporter of interest to strike the sample. An emission filter can be employed (e.g., coated upon or fixed to regions of the upper plate) that allows the fluorescence from the sample to pass to a detector and blocks stray light from the light source or interfering components in the sample.
In an exemplary arrangement, as depicted in the sectional view of FIG. 2, a lower glass or plastic plate 28 is provided with spaced-apart etched channels 14. While each channel 4 is shown having vertical sidewalls 14 a and a flat bottom or floor region 14 b, which meet at ninety-degree angles, other channel geometries can be employed (see, e.g., FIG. 3). The sidewalls 14 a of the channels have bandpass characteristics that permit passage only of the excitation (laser) beam 48 through the device (e.g., a coating material, as at 52, applied to the channel sidewalls 14 a and, optionally, the floor regions 14 b; or micro-optical elements 54 attached on each sidewall 14 a (see FIG. 2B); or the whole lower etched plate (including the channels in their entireties) can be coated with a bandpass coating permitting only excitation light to pass (see FIG. 2C)). An upper or cover plate 26, overlaid over the channels 14, can be provided with bandpass characteristics that let only sample emission (e.g., fluorescence) to pass through (and not excitation beam wavelengths). For example, a bandpass coating material, as at 56 (FIG. 2A), can be applied to a face or face regions of the upper plate 26 that confront the lower plate 28 and channels 14, at least along regions along the detection zone. Or, a micro-optical bandpass element, such as element 55 in FIG. 2B, can be attached to the side of upper plate 26 confronting lower plate 28. By these or similar arrangements, excitation light is permitted to pass laterally through the device in a side-entry, on-channel configuration (with light of other bandwidths being substantially excluded), while fluorescent emission from sample zones is permitted to pass out of the channel to a detector (with light of other bandwidths being substantially excluded).
A number of commercial entities produce a wide range of coating technology products that can find use with the teachings herein, including bandpass filters, beamsplitters, reflectors and collectors, sputtered metals, etc. (For example: Seoul Precision Optics Co.; GM Vacuum, A Division of Navitar Coating Labs; Optical Coating Laboratory, Inc.; and Guernsey Coating Laboratories Inc.).
Optical elements and coatings useful in connection with the present teachings are described, for example, in U.S. Pat. Nos. 3,466,120; 6,112,005; 5,872,655; 4,663,557; 6,100,541; each of which is expressly incorporated herein by reference.
According to various embodiments, an excitation beam of light is directed toward the array from each lateral side of the device. For example, two lasers can be employed, one on each side of the device. Or, as depicted in FIG. 4, a single laser source 34 can be employed in combination with appropriate beam splitting and directing optics, such as at 60 a-60 f, so that light enters the array from each lateral side of the device.
It should be noted that, in some instances, one of skill in the art may choose to use less than all of the channels provided by the device of the invention to conduct separations. For instance, the left- and right-endmost channels might not be loaded with samples, while the remaining channels are so loaded; or, every other channel might be utilized, with the intervening channels remaining unused.
Various embodiments of a channel device include a substrate, with a plurality of channels formed in the substrate. Each channel includes an inlet end and an outlet end. The channels are disposed in spaced relation relative to one another, with each adjacent pair of channels being separated by a respective portion of the substrate that includes at least a region that is transparent. An excitation-beam source is adapted to direct a beam of coherent light along a beam path that intersects each of the channels at a region between the inlet and outlet ends, and further intersects the transparent region of the substrate separating adjacent pairs of channels.
According to certain embodiments, each channel includes opposed sidewalls with portions that are substantially parallel to one another. The parallel portions are at least in part transparent, and the beam path extends through the transparent, parallel portions. Such construction can be useful to avoid or reduce loss of light intensity as the beam travels through the device, from channel to channel.
All publications and patent applications referred to herein are hereby incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.
Those having ordinary skill in the art will understand that modifications are possible in the above embodiments without departing from the teachings thereof. All such modifications are encompassed within the following claims.
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Posts with tag "Gary Busey"
Gary Busey Is to Blame for Crazy Celebrity Baby NamesPosted May 12th 2011 at 11:35 AM by AOL TV Editors
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It has become a staple in Hollywood for celebrities to name their offspring something bizarre. Alicia Silverstone is the latest star to give a unique name to her brand new bundle of joy. Bear Blu is the chosen name for her newborn son and we can't help but wonder where does she, and the numerous other celebrities, come up with these names? 'Jimmy Kimmel Live' (Weeknights, 12AM ET on ABC) had th...
Gary Busey's 1-Year-Old Son HospitalizedPosted Apr 20th 2011 at 03:00 AM by Christo Varghese
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It's been rough week for Gary Busey. On Sunday, Busey got eliminated on 'The Celebrity Apprentice' and now it's reported that his 1-year-old son Luke has been hospitalized. TMZ is reporting that Luke was hospitalized at Cedars-Sinai Medical Center in Los Angeles for an undisclosed "childhood illness." The actor's rep did not get into any specifics but said that Luke was expected to be bac...
Video: Gary Busey Finally Gets Fired on 'Celebrity Apprentice'Posted Apr 18th 2011 at 05:30 AM by AOL TV Editors
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It's been no secret that Meat Loaf has been struggling with various aspects of his time on 'Celebrity Apprentice'. But the greatest source of frustration for him has been Gary Busey. Almost from the beginning, the two of them have not gotten along. It even culminated in a largely undeserved and disturbing expletive-filled freak-out by Meat Loaf toward Gary Busey over art supplies. This week,...
Video: Meat Loaf has an Epic Meltdown on 'Celebrity Apprentice'Posted Apr 4th 2011 at 05:50 AM by AOL TV Editors
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Hey, Gary Busey. Bet you didn't see that one coming, did you? Neither did anyone else, but Meat Loaf certainly had the mother of all meltdowns on 'Celebrity Apprentice' this week. And it was all over art supplies. See, when you can't find your art supplies, you have two options. You can either start looking around for them and asking if anyone's seen them, or you can do what Meat Loaf did. W...
The Original 'Winner': Gary Busey's Most Outrageous MomentsPosted Mar 7th 2011 at 05:00 PM by Kiki Von Glinow
In what should be sold as a DVD box set of the prolific Gary Busey's most enlightening "Buseyisms," he proves that there is a fine line between genius and insanity. Unfortunately, Busey most often finds himself falling off the deep end of the latter. From directing his own interviews, to giving motivational speeches to littering the streets of New York with pepperoni, forget Charlie Sheen, vict...
Gary Busey, Steffanie Sampson Welcome Baby BoyPosted Feb 23rd 2010 at 04:40 PM by PopEater Staff
- Filed under:
- Celebrity Baby -
- Good News -
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Gary Busey and girlfriend Steffanie Sampson welcomed a baby boy this morning, Entertainment Tonight is exclusively reporting. Luke Samson Busey was delivered at 6:40AM and weighed six pounds, seven ounces. The couple announced Sampson's pregnancy early last December. In a statement, Busey and Sampson said that the delivery "feels like a dream. We feel so blessed and are overwhelmed with happ...
Gary Busey, Girlfriend Expecting BoyPosted Dec 10th 2009 at 06:00 AM by PopEater Staff
- Filed under:
- Celebrity Baby -
- Movie News
Gary Busey and Steffanie Sampson will welcome a child together next year. The actor's girlfriend is seven months pregnant with a boy, his agent tells PEOPLE. Just this week he told Moviefone that he's putting his career on the shelf for now, as "working on the birth of this baby in two months" was "all that matters." "He feels awesome about it, he's excited and thrilled and can't wait," his ...
Ten Celebrities Who Need To Join TwitterPosted Nov 8th 2009 at 09:05 AM by Amber James
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- Music News -
- TV News -
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There comes a time when most celebrities should have a Twitter account, and the time is now. Honestly, if Rabbi Shmuley Boteach has an active account before you, it's time to get with the times. Yes, celebs are busy, but we have a wish list here at PopEater of stars we keep hoping will join the social media blog. See who made the list >> ...
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|160 Gram
|Light brown sugar
|300 Gram (Measured Without Packing)
|1⁄4 Teaspoon
|Finely ground blanched almonds
|50 Gram (1 Cup)
|6 Tablespoon
|All purpose flour
|400 Gram (3 Cups)
|Ground cinnamon
|1 Teaspoon
|1⁄4 Teaspoon
|1⁄4 Teaspoon
|1⁄4 Teaspoon
|1 Cup (16 tbs)
Cream butter and light brown sugar together.
Beat in salt, ground almonds, egg, and 3 tablespoons milk.
Add flour and spices and enough of remaining milk to form a stiff dough.
You may need to knead the dough to work in all the flour.
Refrigerate dough for 30 minutes.
If using Spekulatius or any other cookie molds, dust each with flour or cornstarch.
Press enough dough firmly into each mold to pick-up pattern.
Trim excess dough around edges of mold with knife.
Molded cookies will be somewhat thick, about 1/4 inch (3/4 cm) or so.
Unmold cookies onto greased baking sheets, leaving at least 1 inch (2 1/2 cm) of space between each cookie.
Brush cookies lightly with milk.
Bake at 400°F. (200°C) for 10 to 12 minutes, or until lightly browned.
For cookie cutter cookies, roll dough 3/8 inch (1 cm) thick on lightly floured board.
Cut into desired shapes and place on greased baking sheets, leaving 1 inch (2 1/2 cm) of space between each cookie.
Brush with milk and bake.
Cookie cutter cookies will bake a little faster because of their more uniform size and slightly thinner dough.


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posted at 7/28/2011 5:50 PM EDTPats: I think I need a shot of something now. That just made me want to cry myself to sleep. LOL! But on re-read...maybe I should edit that statement. *facepalm*
Re: Sex offenders
posted at 7/28/2011 5:52 PM EDTlol
posted at 7/29/2011 11:50 AM EDTCheck this out on our latest registree - Dennis T. His convictions were in MO where he served 12 years as a child molester and apparently, on his release, didn't feel that he should have to register with the board. Now he lives in Malden, MA. Hooray! (I didn't bother to register for the full article, but this is definitely our newest resident in Ward 6): http://goliath.ecnext.com/coms2/gi_0199-10483470/Missouri-sex-offender-doesnAEt-want.html


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Orders must be placed in the US, they cannot be placed internationally.
Browse and select the photo you want to have printed on your gift card.
We review each image submitted. While we accept most images, we will not accept images that contain any of the following:
We are thoroughly delighted with Blue Diamond! It was easy to make an appointment and the crew was courteous, conscious, and delivered spectacular results....


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34 year old man | Sarasota, FL, USA | Seeking women 30-43 within 55 miles
Active within 3 days
Fun Loving Looking For The Same
I am a good, honest, reliable man. I have a good job. I have a great supporting cast of friends and family. I am looking for the same. Someone who has their feet on the ground, has a good job, loves to have fun, loves to be spontaneous, is looking for a relationship, and is kind and caring. I am looking to travel the world and enjoy all that life has to offer.
Camping , Coffee and conversation , Cooking , Dining out , Fishing/Hunting , Hobbies and crafts , Movies/Videos , Museums and art , Music and concerts , Exploring new areas , Nightclubs/Dancing , Performing arts , Playing cards , Playing sports , Political interests , Shopping/Antiques , Travel/Sightseeing , Volunteering , Watching sports , Wine tasting
Baseball, Basketball, Billiards / Pool, Bowling, Cycling, Dancing, Football, Golf, Hockey, Swimming, Walking / Hiking, Weights / Machines
I exercise 5 or more times per week.
I like Birds, Cats, Dogs, Exotic Pets, Fish, Horses, Other
Not a fan of spiders or snakes...
1. Downtown Sarasota 2. South Beach Miami 3. Biloxi MS 4. Erie, Pa/Pittsburgh Pa 5. St Pete===Tropicana Field 6. Orlando 7. The Florida Keys 8. Any Tropical Islands in the Caribbean 9. New Orleans
Jones International University, Centennial, CO
Music: slow jams such as Boyz II Men, 90s Rap, Hip Hop, Alternative, Country, Reggae, Reggaeton, Latin Music TV: Man vs Wild, Deadliest Catch,, NCIS, Dog The Bounty Hunter, Paula Dean, Steven Segal: Lawman, Cosby Show, Golden Girls, The First 48,
For Fun: beach, swimming, hot tubs, boating, fine dining, cooking, movies, theater, travel, cruises, casinos, concerts, live music, night clubs, family outings, mini-golf, real golf, board games, theme parks
I read tons of books due to being a teacher, but the last pleasure book I read was Roberto Clemente...It was a biography tribute.
Z{{Cta}} {{#if HasMoreInfo}} ?{{CtaSecondary}} {{/if}}
|What he is looking for
|5'8" (172cms)
|5'0" (152cms) to 5'9" (175cms)
|Body type:
|Athletic and toned
|Slender, Big and beautiful, About average, Curvy, Athletic and toned, Full-figured, A few extra pounds, Stocky
|Black, Blue, Brown, Grey, Green, Hazel
|Dark brown
|Auburn / Red, Black, Light brown, Dark brown, Blonde, Dark blonde, Platinum
|No Way
|No Way, Yes, but trying to quit
|Social Drinker
|Social Drinker, Regularly, Moderately
|I'll tell you later
|Administrative / Secretarial, Artistic / Creative / Performance, Executive / Management, Financial / Accounting / Real Estate, Labor / Construction, Legal, Medical / Dental / Veterinary / Fitness, Political / Govt / Civil Service / Military, Retail / Food services, Retired, Sales / Marketing, Self-Employed / Entrepreneur, Student, Education / Teacher / Professor, Technical / Science / Computers / Engineering, Travel / Hospitality / Transportation, Other profession, Nonprofit / Volunteer / Activist, Law enforcement / Security / Military, Fashion / Model / Beauty, Architecture / Interior design
I am middle school social studies teacher, I also have a master's in administration. I have been teaching 11 years and love my job in the fact that I can influence 100s of children each and every day.
|$35,001 to $50,000
|Less Than $25,000, $25,001 to $35,000, $35,001 to $50,000, $50,001 to $75,000, $75,001 to $100,000, $100,001 to $150,000, $150,001+
|Never Married, Widow / Widower, Currently Separated, Divorced
|Have kids:
|Yes, and they sometimes live at home, No, Yes, and they live away from home
|Wants kids:
|Probably not
|Definitely, Someday, Not sure, Probably not, No, No, but it's OK if my partner has kids
|White / Caucasian
|Asian, Black / African descent, East Indian, Latino / Hispanic, Middle Eastern, Native American, Pacific Islander, White / Caucasian, Other
German Irish Native American
|Christian / Catholic
|Agnostic, Buddhist / Taoist, Christian / Catholic, Christian / LDS, Christian / Protestant, Hindu, Jewish, Muslim / Islam, Spiritual but not religious, Other, Christian / Other
|Graduate degree
|High school, Some college, Associates degree, Bachelors degree, Graduate degree, PhD / Post Doctoral


text:
meadowbrook insurance group
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BOARD MEMBERS AFFILIATED WITH Robert H. Naftaly CPA*
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Posted on 29 April 2014
- 19:30
Jan Vertonghen and Etienne Capoue both played a full part in training having returned to action after injury for our Under-21s against Reading on Sunday. Alex Pritchard is also back from his loan at Swindon Town.
Below - Les Ferdinand with Etienne Capoue, Emmanuel Adebayor and Nacer Chadli...
9 April 2015
20 January 2015
30 December 2014
View mobile site
© Tottenham Hotspur Ltd 2014. All rights reserved.
All photography copyright Action Images plc (unless otherwise stated)


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Show Pony Magazine – Issue 2
In Issue 2 of Show Pony Magazine, we celebrate the ingenuity and independence of the American Entrepreneur.Show Pony Magazine is an online community that features extraordinary individuals, artisans and independent business owners. We encourage the creative community, provide education, support small business and aspire to connect with like-minded individuals.
SHOW PONY MAGAZINE ndless E Summer Camp Wandawega has been the host of beloved childhood memories for decades. Rescued on the cusp of fading away, creative entrepreneurs Tereasa Surratt and David Hernandez have given the camp a new lease on life. Photography and Story by Rebecca Hill 122


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Jerez and Cadiz Day Trip from Seville
- Day trip to Jerez and Cadiz from Seville
- Amazing equestrian ballet at the Royal Andalusian School of Equestrian Art in Jerez
- Wine tasting
- Free time in El Puerto de Santa Maria
- Boat trip to Cadiz
- Panoramic tour of Cadiz
- Expert Spanish- and English-speaking guide
- Pick-up and drop-off from selected Seville hotels
Recent Photos of This Tour
What You Can Expect
After the show you'll continue by bus to El Puerto de Santa Maria, where you'll have free time for lunch (additional cost). Later, join a boat trip (or coach, subject to sea conditions) from Puerto de Santa Maria to Cadiz, the oldest populated city in Europe. You'll take a panoramic bus tour of this ancient town before returning to Seville by coach.
Customer Reviews
December 2014
Well this was the best experience of my stay . Our guide was very knowledgeable and spoke perfect English. The whole day ran smoothly, the horses were beautiful just ap shame we could not take phtograghs, even after the show. Enjoyed the whole day immensely , thank you
December 2014
The guide was very knowledgable and very personable. Perfect itinerary led by a perfect guide and driver. Especially liked that it was a small bus with only a few fellow travelers.


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Time = life; therefore, waste your time and waste of your life, or master your time and master your life.
– Alan Lakein
More Quotes
- No life is a waste. The only time we waste is the time we spend thinking we are alone. – Mitch Albom
- We cannot waste time. We can only waste ourselves. – George M. Adams
- Don't waste your time on a guy that isn't willing to waste his time on you. – Anonymous
- The 'Net is a waste of time, and that's exactly what's right about it. – William Gibson
- Doing a thing well is often a waste of time. – Robert Byrne


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Big voices lift San Diego Opera's dark 'Il Trovatore'
When San Diego Opera presented "Samson and Delilah" last month, it was epic in scope -- colossal sets, giant chorus numbers, big dance scenes and a huge star, Denyce Graves.
By contrast, the company's new production of Verdi's "Il Trovatore" goes in the opposite direction, with minimalist sets, 50 fewer people onstage, no dance scenes and a cast of singers who are mostly new to San Diego. Yet the production is still epic in a way that counts -- it's packed with big, beautiful voices that really make Verdi's tuneful score sing.
Big doesn't always mean better in the vocal department, but the "Il Trovatore" cast features singers with exceptional vocal power, combined with nuance, flexibility and finesse. Nowhere is this more apparent than in the San Diego Opera premiere of soprano Paoletta Marrocu as the doomed heroine Leonora. She's a fine actress, and her rendering of the heartbreaking third-act aria "D'amor sull'ali rosee," from its roof-raising high notes to its almost-whispered passages, is the highlight of the evening.
And as the vengeful Gypsy Azucena, Marianne Cornetti is world-class, both vocally and in the emotion she brings to the role.
"Il Trovatore" is known for its hit-packed score, but its story -- set in war-torn Spain in the 15th century -- is a little convoluted. In a nutshell, a young noblewoman named Leonora is courted by the Count Di Luna, but Leonora loves Manrico, a mysterious troubadour (il trovatore) who sings by her window each night. Jealous of Manrico, who is also a leader in the peasant resistance, Di Luna plots to capture and kill him. Adding to the mystery of Manrico is his "mother," the Gypsy Azucena, who vows to avenge the death of her mother, whom Di Luna's father burned at the stake many years before.
British director Stephen Lawless makes his San Diego debut with a sparing "Il Trovatore" production that's been produced around the Globe. The set is simple -- tall black walls (with vertical sliding panels that serve as doorways) that move on, off and around the stage in an ever-shifting variety of patterns. Swords are everywhere in the set -- sprouting from the floor, hanging from the ceiling and waved about by the cast in a number of scenes -- and noisy jets of fire occasionally burst from the floor.
Except for some opening-night clunks and scrapes, and lighting that was almost unrelentingly gloomy, the sets worked quite effectively to portray a variety of locations and reflect the opera's dark subject matter, while virtually eliminating the need for lengthy scene changes.
Lawless has reinterpreted some scenes, moving the singers around in nontraditional ways that sometimes work and sometimes don't. One idea that does is his blocking of the chorus, first in the famed "anvil chorus" (where the Gypsies crawl like rats through small holes in the walls, then scale the walls for dramatic effect), then in the third-act opener, where the soldiers chorus performs "Or co'dadi ma fra poco" while they perform a synchronized sword-training drill, en masse.


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The U.S. Army has awarded General Dynamics C4 Systems a contract with a potential value of $50 million to provide the service's Land Warrior program with engineering and logistics support. The contract covers a one-year base valued at $19 million, with options for two additional years.
General Dynamics
The U.S. Navy's Naval Surface Warfare Center has awarded General Dynamics Information Technology a contract for hull, mechanical, electrical and electronic systems alterations installation team support. General Dynamics is one of six firms awarded the five-year, indefinite delivery/indefinite quantity contract with a potential value of $475 million.
The U.S. Navy has awarded General Dynamics Electric Boat $61.8 million to extend two existing contracts for submarine modernization, maintenance and reactor plant planning yard work.
General Dynamics has been awarded a $152,818,728 cost-plus-incentive fee contract for fiscal year 2010 and fiscal year 2011 for production and deployed systems support for the United States and United Kingdom SSBN fire control system and SSGN Attack Weapon Control System, including training and support equipment.
General Dynamics has been awarded a five-year, $140 million contract from the U.S. Coast Guard to provide information technology (IT) enterprise engineering, operations and maintenance support services for both classified and unclassified elements of its computing environment. The contract work will cover areas of the Coast Guard's land-based and mobile telecommunications, electronics and information systems.
General Dynamics Advanced Information Systems has been awarded a $152.8 million contract by the U.S. Navy for fiscal years 2010 and 2011 production and deployed-systems support for the United States and United Kingdom SSBN fire control system (FCS) and the SSGN Attack Weapon Control System (AWCS).
General Dynamics Awarded $86 Million for Sustainment of Marine Corps' Mobile Combat Operations Centers
General Dynamics Information Technology has been awarded an $86.2 million contract to provide field-level sustainment services and technical support for equipment and networks for the U.S. Marine Corps' critical Combat Operations Center program.
General Dynamics Land Systems has been awarded a $24 million contract to supply Commanders Remote Operated Weapons Stations, version two (CROWS II) kits to the U.S. Army TACOM Lifecycle Management Command. CROWS II is a remote-controlled weapon system that provides soldiers the ability to acquire and engage targets from inside the protection of an armored vehicle.
General Dynamics C4 Systems has received a $37 million contract to supply the U.S. Air Force with identification friend or foe (IFF) Mode 5 (KIV-78) cryptographic modules and spare parts.
The U.S. Navy has awarded General Dynamics Electric Boat a $31 million contract to plan and perform maintenance and modernization work on U.S. Trident-class and U.K. Vanguard-class submarines.
General Dynamics Information Technology is being awarded a $10 million contract for the Trans Regional Web Initiative in support of U.S. Special Operations Command (SOCOM) Joint Military Information Support Command.
The U.S. Army TACOM has awarded General Dynamics Armament and Technical Products a $100 million contract for the production of M2 flex machine guns.
General Dynamics Information Technology has received a $22 million contract for design and development of one release of Naval Integrated Tactical Environmental System-Next Generation (NITES-Next) software.
The U.S Navy has awarded General Dynamics' subsidiary, Electric Boat, a $42.1 million contract to plan and perform routine maintenance and modernization work on USS Springfield (SSN-761), a Los Angeles-class attack submarine.
General Dynamics Electric Boat Corporation is being awarded a $6 million modification to a previously awarded contract for Nuclear Regional Maintenance Department tasks in support of operational nuclear submarines.
General Dynamics C4 Systems has received a $17 million contract modification from the U.S. Marine Corps to build nine Combat Operations Center (COC), Capability Set III systems.
General Dynamics Advanced Information Systems is being awarded a $21 million contract for engineering services in support of the AN/BYG-1 Tactical Control System.
General Dynamics Amphibious Systems has been awarded a $6 million contract for two MK 46 Mod 2 gun weapon systems.
General Dynamics Land Systems has been awarded a contract by the U.S. Army TACOM to reset 330 Stryker infantry combat vehicles. The contract has a maximum potential value of $55.2 million.
The U.S. TACOM Lifecycle Management Command has awarded General Dynamics Land Systems a $45 million contract for the Egyptian tank coproduction program.


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Udivitel'nye Podvigi
English Title: Astonishing DeedsUdivitel'nye Podvigi (Astonishing Deeds) (1976) Theatrical Cartoon by Dave Koch
This page has been viewed 6 times this month, and 183 times total.
- Theatrical ShortShort
- Traditional Animation
- Ekran
- Cartoon Characters: Svistulkin.
- Voice Actors: Vyacheslav Nevinny, B. Zakharova, Liya Akhedgakova, Ludmila Ivanova.
- Directed By Boris Ardov.
- Animated By Sergei Olifirenko, A. Degtyarev, Olga Degtyareva, Alla Grishko.
- Written By Nikolai Nossov, Valeria Konovalova.
- Music: Igor Yefremov.
- Editor: Galina Drobinina.
- Originally Released in December of 1976 (Estimate).
- Originally Released Theatrically.
- Running Time: 16:09 minutes.
- Color
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36 CFR 330.8 - Annual report.
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(RCS-DAEN-CWO-53) The Division Engineer will submit a consolidated annual report to reach HQDA (DAEN-CWO-R) WASH DC 20314 not later than 30 October. This requirement expires 30 October 1979. The report will contain the following:
Title 36 published on 2014-07-01.
No entries appear in the Federal Register after this date, for 36 CFR Part 330.


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Brookford Farm is a 650-acre diversified farm that grows vegetables, raises dairy cows, pigs, chickens. We produce an assortment of cheeses and yogurts, grows grain, and has a small bakery on site.Located in Canterbury, NH on the banks of the Merrimack River and located 15 minutes north of Concord, NH, Brookford Farm uses organic methods and is in the process of restoring this former sod farm into productive sustainable agricultural use. Brookford Farm produces a wide variety of meets cheeses and produce for a 250-member year round CSA, weekly farmer's markets, and a farm store on site. For more information about the farm and our mission, please visit: http://brookfordfarm.com
2) Vegetable Apprenticeships
Apprentices will learn many aspects of operating an organic vegetable farm
through various hands-on tasks including greenhouse work, tractor work,
harvesting, and distributing and marketing produce. Throughout the growing
season, apprentices will work closely with the Vegetable Manager through frequent
field walks, check-in meetings, and discussions about best farm management
practices and strategies.
We seek individuals that have a strong interest in sustainable agriculture and
in working on a fast-paced production farm. Previous farming experience is
a plus, but not required. We are looking for individuals with a good sense
of humor who enjoy working hard, are excited to learn, and are responsible and
flexible. You must be able to clearly follow and give directions, not be
afraid to ask questions and make mistakes, and communicate well with others.
You must be able to perform physically demanding and repetitive tasks, work in
all kinds of weather, and be able to safely lift 50 lbs.
Our farm apprenticeship runs full-time from April 1st – November 16th, with possibilities of staying longer for part-time work. While working for the entire season is preferred, it is not required. There is however a minimum requirement of 20 weeks. The work schedule generally runs from Monday through Friday and the hours are 7am – 5pm with an hour for lunch for the majority of the season. In addition there will be rotating work hours on weekends to water and work on irrigation. There may also be extended hours in the more intense part of the season.
Apprentices are paid $225 per week and produce from the farm. Housing is provided, in a shared house for farm apprentices.
To Apply
Please send in a cover letter, resume, and two references to firstname.lastname@example.com Applications will be reviewed on an ongoing basis until filled so please send in your application ASAP. Please contact us if you have any questions.
Brookford Farm is a participating mentor farm in the North American Biodynamic Apprenticeship Program (NABDAP). For more details, please visit the farm's profile in our NABDAP program pages.


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Product Specs
Product Description
Present these lovely cones filled with our Jordan almonds as favors. Cones are available in white or ivory color with an embossed floral design. Also available in white quilted, metallic silver, gold, chocolate brown and clear. Measures 6 inches long.
Coordinate with the color scheme of the birtday party. Choose from one of dozens of colors in our Midori ribbon collection. Or add a custom tag for that personalized touch.


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In This Section
Journal Forum
Clean Audio for TV broadcast: An Object-Based Approach for Hearing-Impaired Viewers - April 2015
2 comments
Audibility of a CD-Standard A/DA/A Loop Inserted into High-Resolution Audio Playback - September 2007
13 comments
Sound Board: Food for Thought, Aesthetics in Orchestra Recording - April 2015
2 comments
AES Conference Papers Forum
Low Frequency Room Excitation using Distributed Mode Loudspeakers
Conventional pistonic loudspeakers excite the modes of an enclosed sound field in such a way as to introduce modal artefacts which may be problematic for listeners to high-quality reproduced sound [1]. Their amelioration may involve the use of highly space-consumptive passive absorptive devices or active control techniques [eg 2,3,4]. Other approaches have concentrated on the design of the driver used to excite the room. Distributed sources ranging from the dipole [5] to more complex configurations [6] may be expected to interact with the room eigenvectors in a complicated manner which may be optimised in terms of the spatial and frequency-domain variance of the soundfield. Recent interest in distributed sources has centred on the Distributed Mode Loudspeaker (DML), and this paper reports an investigation into the interaction of DMLs with modal soundfields. It is shown that large DMLs may be expected to modify the low-frequency soundfield. Producing useful low-frequency control remains difficult but may be achieved in some circumstances.
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Originally posted by NoRegretsEver
reply to post by inverslyproportional
I havent heard of an irok-z in sooooo long, they were popular in NYC in the late 80's and early 90's, its still considered a classic, and you were the man/woman if you had one.
Peace, NRE.
I love to fix things, carpentry wise, but am terrified to mess with electricity, and hate reading directions (yeah I know).
Originally posted by NoRegretsEver
reply to post by cody599
Thats awesome, when I was a kid I was into wood working mostly with lathes, and I loved it, I went to class for carpentry and since I was the only girl there, I was introduced to the lathe, and LOVED it, I have six stitches in my hand because I really didnt pay attention in the beginning to how sharp the chisels were
Peace, NRE.


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Scouts all set to collect cash at the Pirelli Stadium
Posted: April 23, 2014|
DOZENS of scouts will be out in force before, during and after Burton Albion's final home game of the season to raise the profile of the organisation and raise some cash to boost its coffers.
Bosses at the League Two club revealed that the Pirelli Stadium would be hosting members of the 11th Burton on Trent Scout Group, also known as 'The Foresters', at Sunday's game against Chesterfield.
The group is hoping to raise money to pay for a trip to Belgium in the summer in a bid to commemorate the start of the First World War.
Angela Burns, group scout leader, revealed why the group will be heading to the Pirelli at the weekend.
She said: "We would like to thank everyone in advance who will drop some change into our buckets.
"We as a group and provide weekly activities to around 135 young people across the area.
"We are currently raising funds to allow a group of 34 scouts and leaders to visit Belgium this summer.
"We were looking for a venue for our summer camp and, with the anniversary of the start of the First World War this year, we decided to go and visit Belgium and take in some of the history of the war.
"We have planned to visit some of the battlegrounds and war graves and are paying particular attention to those young men that left our area to go and fight for their country.
"We will pay our respects to as many soldiers that are buried or remembered there as well as we can.
"We try to make all trips affordable to everyone and the kind generosity of fans will help us to achieve our goal."
The scouts will be out and about at the stadium in various locations before and after the game.
More information is available by heading online to www.forestersscouts.org