text:
An apparatus is provided for characterizing cells in a biological liquid sample and operating according to the Coulter principle. The apparatus has a disposable sensor unit and a docking station for removably receiving the sensor unit. The sensor unit housing contains electrodes in each of two chambers separated by a wall containing an orifice. The housing is adapted to contain and retain for disposal liquid pumped through the orifice during particle characterization operations.
1. Apparatus for characterizing cells in a biological liquid sample, comprising a sensor unit and a docking station for removably receiving the sensor unit,
the sensor unit adapted for collection and storage of the liquid sample before and after measurement, and comprising:
a housing with
a first chamber for holding the biological liquid sample before measurement,
a collection chamber for collection of the liquid sample after measurement and having an inlet/outlet for connection to a source of positive or negative gas pressure,
a wall separating the first chamber and the collection chamber and containing an orifice for the passage of the cells from the first chamber to the collection chamber, and
a first electrode in the first chamber and a second electrode in the collection chamber, each electrode being electrically connected to a respective terminal member accessible from the exterior of the sensor unit,
the docking station comprising
a port for connection with the source of positive or negative gas pressure and forming a gas connection with the inlet/outlet when the sensor unit is received in the docking station, and
electrical connector members for connecting to the respective terminal members when the sensor unit is received in the docking station.
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9. A sensor unit for characterizing cells suspended in a biological liquid sample and adapted to be removably received in a docking station, comprising
a housing adapted for collection and storage of the liquid sample before and after measurement, and including
a first chamber for holding the biological liquid sample before measurement,
a collection chamber for collection of the liquid sample after measurement and having an inlet/outlet for connection to a source of positive or negative gas pressure,
a wall separating the first chamber and the collection chamber and containing an orifice for the passage of the cells from the first chamber to the collection chamber, and
a first electrode in the first chamber and a second electrode in the collection chamber, each electrode being electrically connected to a respective terminal member accessible from the exterior of the sensor unit, in such a way that,
when the sensor unit is received in the docking station, the inlet/outlet is operationally connected with a port in the docking station for connection with the source of positive or negative gas pressure and electrical connector members are operationally connected to the respective terminal members.
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11. Sensor unit according to
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14. Sensor unit according to
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16. Sensor unit according to
17. A method of operating a particle characterization apparatus comprising:
a sensor unit that is demountable from the apparatus and contains a first chamber for holding a biological liquid sample, and a collection chamber separated by a wall containing an orifice for passage of cells from the first chamber to the collection chamber that initially contains a volume of air and has a connection to an air pump for pumping liquid through the orifice into the collection chamber, and further comprising a first electrode in the first chamber and a second electrode in the collection chamber, each electrode being electrically connected to a respective terminal member accessible from the exterior of the sensor unit,
the apparatus further comprising electrical connector members that are operationally connected to the respective terminal members when the sensor unit is received in the docking station, the method comprising
passing a biological liquid sample containing cells from the first chamber, through the orifice, and into the collection chamber,
making cell characterizing measurements for a period in such a way that the liquid does not flow out of the collection chamber,
disconnecting the sensor unit from the apparatus with all of the liquid sample that has passed into the collection chamber, and
discarding the sensor unit.
This application is the national phase under 35 U.S.C. � 371 of PCT International Application No. PCT/EP00/07537 which has an International filing date of Aug. 3, 2000, which designated the United States of America and was published in English.
The present invention relates to particle characterisation apparatus in which particles suspended in a liquid are passed through an orifice, in principle in single file, to enable the characterisation of the particles, for instance by Coulter counting.
By aspiration of particles through a small orifice the particles can be characterised with respect to size, concentration and conductivity by the use of an electrical impedance technique, widely known as the Coulter sizing (see V. Kachel, �Electrical Resistance Pulse Sizing: Coulter Sizing�, Flow Cytometry and Sorting, Second Edition, pp. 45-80, 1990 Wiley-Liss).
Different methods involving optical and electrical ways of sizing particles are frequently used for diagnostics and are employed in sophisticated instruments, such as Flow cytometers and Coulter sizing equipment. Until now there have been only a few instruments available for particle counting and sizing, which still needs fairly expensive equipment that requires trained personnel to perform the measurements. There have been several groups working on micro-machined optical flow cytometers (A. Y. Fu et al, �A micro-fabricated fluorescence-activated cell sorter�, Nature Biotechnology, Vol. 17, November 1999, pp. 1109-1111), which could be used as cheap portable instruments. However, these instruments still require precision fluidic dispensing, a laser and a sensitive detection system. As an alternative, the much simpler Coulter method (explained in next section) can be employed. Such an instrument would be limited to counting and sizing of cells but still be an attractive tool for a wide variety of applications, e.g. milk quality control and blood cell counting.
Counting and sizing of particles by the Coulter principle is an internationally respected method that is being used in most haematology-analysers and particle counting equipment (see FIG. 3). The method is based on measurable changes in the electrical impedance produced by non-conductive particles in an electrolyte. A small opening called the �aperture or orifice�, connects two electrically isolated chambers, where electrodes has been provided to contact the electrolyte. The orifice applies an restriction to the electrical path, whereby a sensing zone is established through which the particles are aspirated. In the sensing zone each particle will give rise to a displacement of the surrounding electrolyte, thus blocking part of the current-path and giving rise to a voltage pulse. By this method several thousand particles per second can be characterised with high precision. As opposed to optical methods, this method is not depending on the particle morphology, colour or density.
Conventional instruments utilising the impedance technique are based on a fixed membrane with a precision-machined orifice, which is being maintained by flushing and rinsing the membrane. These procedures require cleaning�and flushing liquids, pumps and control electronics, which complicates and increase the cost of the instrument. In conventional Coulter counters, the orifices are formed in materials such as ruby or sapphire which are chosen for their dimensional stability under the conditions required for their fusion to glass in the construction of sensor elements. These commercially available particle analysers cannot be considered portable, which implies that all equipment are placed centrally in large laboratories. Furthermore, maintenance is obviously required.
Since many laboratories can not use the inflexible standard instruments or can not afford the cost of these, manual methods are still widely used for doing simple counting. These methods are based on a small microscope slide with well defined square markings known as a haemocytometer (see Stevens, �Fundamentals of Clinical Haematology�, W.B. Saunders Company, 1997, ISBN 0-7216-4177-6); The sample is injected in between the gap from the haemocytometer and a cover glass placed on top of the markings. The haemocytometer is placed under a microscope and particles are counted manually. Manual counts are performed until a number of between 100-300 cells has been reached�a method carrying an error of approximately 10%. By electronic counting more than 10,000 cells can be counted within 1 minute, giving an error of 1% or better. Furthermore, timing is critical when counting with haemocytometers, since particles need a little time to settle, but waiting too long will cause an uneven distribution of cells due to evaporation from the open edges. It is not really possible to perform a measurement of cell size with a standard haemocytometer, but with an addition of a high resolution CCD-camera, cross-sectional cell size can be determined (see patent application WO 98/50777).
The cross-sectional area found by 2D images does not necessary relate to the volume of the particles, but depends on the axial symmetry. With the electrical impedance technique it is possible to resolve the particle volume from the measurement. By maintaining a constant current across the orifice, the recorded voltage pulse from particles displacing the electrolyte in the orifice will have a height proportional to the volume of the particle. This is because particles can be considered non-conducting compared to the electrolyte, the electrical field in the centre of the orifice is homogeneous, which is normally the case when the diameter is smaller than the length of the orifice (l/D>1), the particle is to be considered small compared to the diameter of the orifice (d<0.2*D), only one particle passes through at a time and the particles are passed through the orifice in the axial path.
In a first aspect, the present invention provides apparatus for characterising particles suspended in a liquid, said apparatus comprising:
Normally such apparatus is operated so that the flow through said orifice is into the collection chamber.
In some preferred embodiments, the housing is divided into a first chamber and said collection chamber by said wall containing the orifice. The first chamber may then have a breather inlet/outlet for communication with the atmosphere.
Preferably, the sensor unit is designed to be disposable after a single use. It is desirable that after use there is no need to clean the remainder of the apparatus before it can be used in a new assay procedure with a new sensor unit. Accordingly, one should avoid the escape of liquid from the sensor unit and its entry into the docking station. To this end the positioning of the orifice with respect to the breather inlet/outlet, the second chamber inlet/outlet and said particle characterisation device components is preferably such that a volume of liquid sufficient for the desired particle characterisation can be drawn or pumped through the orifice without the liquid passing out of the housing. Generally, it should be possible to pass a volume of liquid which is at least 1 ml, e.g. 5 ml, through the orifice whilst particle characterisation measurements are being made without the liquid overflowing out of the housing.
Preferably, said components of the particle characterisation device associated with the sensor unit include a first electrode in said first chamber and a second electrode in said second chamber, each said electrode being electrically connected to a terminal member accessible from the exterior of said sensor unit, and said means in said docking station for functionally addressing said components comprises respective electrical connector members for connecting to said terminal members. Generally, it is preferred that all necessary electrical and fluid connections to the sensor unit can be established by fitting the sensor unit into the docking station, preferably by a simple push fit.
In one preferred arrangement, the inlet/outlet of the second chamber of the sensor unit leads from a head space in said second chamber through a conduit formed in said housing to a port positioned at the bottom of said housing which co-operates with said port of the docking station. The sensor unit may then be a push fit downwards into the docking station.
Alternatively, the inlet/outlet of the second chamber of the sensor unit leads from a head space in said second chamber through a conduit formed in said housing to a port positioned at the top of said housing which co-operates with said port of the docking station. The sensor unit may then be a push fit upwards into the docking station.
Preferably, said orifice is formed by a process of photo-lithography, e.g. in a silicon wafer. By fabricating the orifice wafers (also known as �count wafers�) for a Coulter counter in such a way, these become much cheaper than orifice wafers for conventional counters on the market. The allows the manufacturing of a sensor unit cheaply integrating the necessary chamber, aperture and electrodes. By doing this, the sensor unit can be used as a disposable unit, which removes the need for the washing and rinsing of the aperture as in conventional instruments. Furthermore it is not necessary to use time to switch the instrument for the size of particles in subject, since this will be specific to the used aperture. The disposable chamber may contain a standard solution (a liquid with known concentration of particles) that can be used for calibrating the sizes and for determination of concentrations. The chambers can be constructed such that the liquids will remain inside the chamber, such that the instrument in which it will be placed will not require rinsing after use.
Preferably, said apparatus includes means for determining the beginning and end of a period during which a predetermined volume of liquid has passed through said orifice.
This may include the provision of a secondary electrode in the first or the second chamber positioned for sensing when liquid in said chamber is at or above a first level, which secondary electrode is connected to a terminal member accessible from the exterior of said sensor unit, and said docking station may then comprise an electrical connector member for connecting to said terminal member of the secondary electrode.
There may be a further secondary electrode in the same chamber as said secondary electrode and positioned for sensing when liquid in said chamber is at or above a second level, which further secondary electrode is connected to a terminal member accessible from the exterior of said sensor unit, and said docking station may comprise an electrical connector member for connecting to said terminal member of the further secondary electrode.
These secondary electrodes may be used for sensing when the level of the liquid is such that the respective secondary electrodes are or are not immersed in the liquid and may therefore serve for determining the beginning and end of a period during which a fixed volume of liquid has passed through the orifice. For instance, particle characterisation may begin when the level of the liquid just falls below the level of a first said secondary electrode and may end when the level of the liquid just falls to below a second of the secondary electrodes, the volume of liquid passing out of the chamber containing the secondary electrodes during this period being defined by the separation of the secondary electrodes. Of course, it is not necessary that the secondary electrodes be in the chamber from which liquid passes through the orifice, they could equally well be in the chamber into which the liquid flows and both secondary electrodes do not need to be in the same chamber.
Where the end point of the passage of a defined volume of liquid is the effective emptying of one chamber to below the level of the orifice, it is preferred that each of the first and second chambers (or at least that chamber from which liquid passes) has a transverse cross sectional area at the level of said orifice which is substantially less than the transverse cross sectional area of said chamber over a substantial part of the height of the chamber above said orifice.
The invention includes a sensor unit for apparatus for characterising particles suspended in a liquid which sensor unit comprises:
Said sensor unit may comprise components of a particle characterisation device for characterising particles passing through said orifice in use, which components are functionally addressable from outside said housing, e.g. a respective electrode in each chamber. Preferably therefore, said components of the particle characterisation device associated with the sensor unit include a first electrode in said first chamber and a second electrode in said second chamber, each said electrode being electrically connected to a terminal member accessible from the exterior of said sensor unit.
Preferably, said orifice is formed by a process comprising photo-lighography, e.g. in a silicon wafer.
Preferably, the sensor unit comprises a secondary electrode in the first or the second chamber positioned for sensing when liquid in said chamber is at or above a first level, which secondary electrode is connected to a terminal member accessible from the exterior of said sensor unit and preferably the sensor unit comprises:
Optionally, each of the first and second chambers has a transverse cross sectional area at the level of said orifice which is substantially less than the transverse cross sectional area of said chamber over a substantial part of the height of the chamber above said orifice.
In a second independent aspect, the invention provides particle characterising apparatus in which particles are characterised as they are passed through an orifice, wherein said orifice is formed by a process comprising photo-lithography, e.g. in a silicon wafer.
An orifice may suitably be formed in a photo-reactive polymer by photo-lithography and subsequent development. Thus a free standing sheet of polymer of the kind used conventionally as a photoresist material may be exposed to light to solubilise a spot to be removed to define an orifice (or to insolubilise the non-spot forming areas) followed by development with solvent to remove material to form the orifice. Normally, a large number of count wafers each containing a respective orifice will be made simultaneously in one sheet. Suitable photoresist polymers are described in e.g. M. Madou �Fundamentals of Microfabrication; CRC Press LLC, 1997, ISBN 0-8493-9451-1. They include AZ-5214E, SU8, polyimides and others.
Alternatively, the photoresist polymer may be used as a protecting layer over a substrate such as silicon or any other etchable material in which the orifice is formed by etching regions exposed by development of the photoresist. If the etched substrate is electrically conducting it may be insulated prior to use by the formation of a suitable insulating layer thereover. The photoresist polymer may be used as such a layer.
Count wafers made lithographically may be used in all forms of apparatus and method according to this invention.
In a third aspect, the invention provides a method of operating particle characterisation apparatus comprising a demountable sensor unit containing a count orifice serving as an inlet to a chamber initially containing a volume of air and having a connection to an air pump whereby air may be pumped to induce flow of sample liquid through said orifice into said chamber, said method comprising passing liquid containing particles through said orifice into said chamber and making particle characterising measurements for a period such that said liquid does not fill said chamber, disconnecting said sensor unit from said apparatus with all of the liquid that has passed into said chamber and discarding said sensor unit.
For use in such a method, the invention includes apparatus for characterising particles suspended in a liquid, said apparatus comprising:
Generally, in all embodiments it is preferred that all components which are wet by the sample in use are disposable and all non-disposable components can be re-used without cleaning.
The invention will be further described and illustrated with reference to the accompanying drawings in which:
The apparatus shown in
The sensor unit comprises a housing 14 having a base wall 16 and a circumambient side wall 18 with a wall 20 rising from the base and separating the housing into two chambers; a first or �open� chamber 22 and a second or �closed� chamber 24. At the upper end of first chamber 22 there is provided a removable membrane seal 26, providing a temporary closure which is removed for use. At the top of the second chamber there is a top wall 28 which closes the chamber. An orifice in the bottom wall of the housing forms an outlet/inlet 30 from which ascends a conduit 32 formed in the wall of the chamber. Conduit 32 ends short of the top wall 28 and so communicates with a head space 34 within said chamber 24.
The dividing wall 20 has a count wafer (not shown) received over a through hole therein and defining a count orifice 36 positioned somewhat above but quite close to the base wall 16. Each chamber has an electrode 38,40 extending from an external terminal 42,44 through the base wall 16 and up to a level within its respective chamber which is somewhat above the level of the count orifice 36.
Particles can be aspirated through the orifice by pressure driven flow in either direction. When a saline or other electrolytic liquid solution is added to the chambers, the two chambers will be electrically isolated from each other except for the route for current flow provided by the passage through the orifice.
The docking station 12 has a cup shaped housing having a base 46 and a circumambient side wall 48. In the base 46 there are respective spring loaded electrical connectors 50,52 for contacting the terminals 42,44 of the sensor unit automatically when the sensor unit is received as a push fit into the docking station. There is also a conduit 54 passing through the base wall 46 aligned with the conduit 32 of the sensor unit. Conduit 54 at its opening into the upper face of the wall 46 has an O-ring seal for forming a gas tight connection with the lower face of the base wall 16 of the sensor unit.
A vacuum pump 58 is connected by a line 60 to the lower end of the conduit 54. In a modification of the apparatus, the vacuum pump 58 can be reversed so as to apply positive gas pressure to the conduit 54.
Schematically indicated at 62 are the further conventional components of a Coulter counter including all the electronic circuitry and display equipment needed for the operation of the apparatus.
The docking station is embodied in a portable instrument into which the sensor unit is a disposable plug-in component. The sensor unit is easily interconnected with the docking station by placing it in the docking station receptacle as illustrated in FIG. 1. The sensor unit is placed in the docking station and fixed with a easy pressure downwards. The electrical contacts to the electrodes are established automatically upon insertion and hermetic sealing to the opening of the suction conduit 32 is automatically established so that suction can be applied by the small air pump 58.
The sensor unit housing can be partly fabricated with cheap mass-production techniques such as polymer injection moulding. Electrodes can be made of small rods of metal or graphite. The orifice can be made very precisely with Silicon photolithography and various etching techniques (wet etching and dry etching, see e.g. �Fundamentals of micro-fabrication�, Marc Madou, CRC Press LLC, ISBN 0-8493-9451-1) and placed in the separating wall. The surface of the silicon wafer containing the orifice may be made electrically insulating by forming a layer of silicon dioxide or silicon nitride over the wafer by baking in an oven in a suitable atmosphere.
Sample liquid with particles in suspension can be applied to the open chamber of the sensor unit. Applying a negative pressure difference to the closed chamber through the conduit 32, will cause the liquid to be sucked through the orifice and into the closed chamber. The suction outlet is led from the head space near the top of the closed chamber, so that only air is sucked out as long as the liquid is below this level. This implies that the docking station is and the remainder of the instrument does not have to be cleaned as would be the case after contact with biological material.
The chambers can be pre-filled with liquids (preferably an electrolyte) and calibration particles for the measurement. This implies that the sensor unit is ready for use before the insertion into the docking station. These particles can be used for calibration of the sizing or as a measure of particle concentration in the sample, if these can be differentiated from the calibration particles. If the concentration of calibration particles is known it is not necessary to know either volume or concentration of the particles in the sample to determine this. The concentration is given by C1=n1/n2*C2 , where C1 is the concentration to be found, C2 is the known concentration of calibration particles, n1 is the number of counted particles from the sample and n2 is the number of counted particles from the calibration standard.
If no calibration particles are present, the concentration of particles in the sample can be determined by control of the volume sucked through the orifice. The apparatus can be configured so that one can determine when a known volume of liquid has been passed through the orifice. This can be done by constructing the container in a way, that would allow the liquid to be removed essentially totally from the open chamber to the closed chamber or at least such that the volume removed by lowering the level in the open chamber to the level of the count orifice is reasonably precise. A way to do this is illustrated in FIG. 7. By sloping the upper surface of the bottom wall 16 of the housing and placing the orifice 136 and electrodes 142, 144 near the bottom of the well so formed in each chamber, it will be possible to suck almost all liquid through the orifice, while contact to the electrodes still remains through electrical connector 150. The volume passed is then determined by the starting volume in the open chamber. The advantage of this method is that the desired applied sample volume may differ in different tests.
As an alternative means of determining the volume passed, one may place secondary electrodes in the chambers that indicate the level of the electrolyte by sensing the presence of conductive media as depicted in FIG. 8. The electrodes 38,40 will be present as in
Finally, since the electrodes 38,40 placed in the chambers are needed in order to count particles, the contact to these will in fact indicate the presence of conductive media. This could be used to indicate the levels of the electrolyte in the two chambers as illustrated in FIG. 9. The electrodes 38,40 are conductive from a certain level and up, which can be established by making the lower part of the electrode non-conductive, e.g. by a non-conductive sleeve 72. When a measured volume of the electrolyte 66 is added to the open chamber 22 of the housing 14 there is no contact between the electrodes. In this situation no pulses will be counted. After a short while, as suction is applied, the level in the closed chamber will reach the conducting portion of the electrode 40 (situation (A)). The counting will start automatically and end when the level in the open chamber is below the level of the non-conductive sleeve of the electrode 38 (situation (B)). This method allows different amounts of sample to be used, and the counting will be of particles in the volume applied less the volume below the conductive part of the electrode 40. This method is somewhat similar to the first method described with reference to
An alternative form of disposable sensor unit and its associated docking station is shown in FIG. 10. It illustrates at least the second and third aspects of the invention. Here the first or open chamber 22 is vestigial and could be omitted entirely. The sensor unit has a housing in the form of a cylindrical tube 14 divided into an upper second chamber 24 and a lower said first chamber 22 by a partition wall 20 containing a count wafer defining a count orifice 36. The count orifice is formed in a silicon wafer attached in the centre of the wall 20. The partition wall 20 can be positioned as low in the tube 14 as desired, even at its extreme lower end.
There are no electrodes forming part of the sensor unit in this embodiment but it could readily be modified such that the electrodes are attached to the tube 14. For instance, one electrode could be located as a tube on the inside of tube 14 with the other being located as a tube on the outside of tube 14 so that tube 14 acts as an insulating spacer between the two electrodes 240 and 238 as shown in FIG. 11. Also shown in
Alternatively, both the sensor unit 10 and the docking station 12 can form part of a disposable assembly, in which case the sensor unit can be permanently fixed in the docking station. The connecting wires of the electrodes can be terminated at connectors such that the whole assembly illustrated in
The methods described can be combined to give the best solution for the final application. The disposable sensor is particularly usable where portable, cheap, simple or flexible equipment is needed, such as in small laboratories, in measurements in the field or as a �point of care� diagnostic tool.
A suitable electrolyte for use in apparatus according to the invention is 0.1 m KCl in filtered water. When sample is applied to the electrolyte, the electrolyte to sample volumes should preferably be higher than 10. Sample preparation should preferably result in between 103 to 105 particles per ml. A vortexing of the sample before and after adding electrolyte is recommended. Particle diameters should be within 10 to 30 percent of the orifice diameter. Volume flow should normally be from 100 μL to 1 ml per minute.
For the measurement a constant electrical current of approximately 1 mA should preferably be applied. The source of electrical current should preferably have a signal to noise ratio (S/N) better than 50000. The response from the electrode should be filtered electronically by a band-pass filter. A second order Chebyscheff filter with centre frequencies around 1 kHz is suggested.
The use of the apparatus described above is further illustrated by the following examples.
A mixture of 6.10 μm and 9.15 μm particles suspended in electrolyte was aspirated through the orifice of the apparatus shown in FIG. 1. The numbers of particles detected and the size of each detected particle were recorded. A bimodal distribution of detected particle size is clearly seen in FIG. 2.
The volumes of the particles are assumed to be proportional to the pulse height, which is only true within a certain interval. Because non-linearity is reproducible, particles that are out of range may be sized by using standard beads with known volumes. As can be seen from
One of the most important, but also rather critical measurements to make with the counter is a concentration determination. In order to get an accurate concentration determination the particle suspension has to be homogeneous, the numbers of counted particles must be high enough and the amount of liquid going through the aperture must be controlled. Furthermore in association with the collection of data on the computer one must consider e.g. sample rate and trigger level. One of the more intrinsic problems is the occurrence of clusters of particles, whereof the most evident are doublets and triplets. These are most frequent when working with non-biological particles such as latex beads. In
The test was run three times with a total volume of the sample of 0.05 ml for each run giving a mean count of 11.384 particles with a standard deviation of 389 (3.4%). Using the correction factor of 1.4 this gives a total corrected concentration of: 319�103�11�103 particles/ml. A small amount of sample was taken from the counting chamber and manually counted in a haemocytometer under a microscope. Two independent manual counting controls resulted in a concentration of: 314�103�25�103 particles/ml. These experiments clearly establish that a good correlation exists between our Coulter counter results and the results from manual counting.
Measurement of blood cells has been performed and the result is shown in FIG. 4. Red blood cells are normally around 5 to 7 μm in diameter and are the most frequent in whole blood, as can be seen on the figure. Less frequent are white blood cells that are 6-10 μm in diameter. The distribution is a Gaussian curve, as it should be expected. Blood counts can be used in clinical diagnostics. It is fairly simple to count erythrocytes, leukocytes and thrombocytes by impedance measurements, which is considered the basic parameters for hematology (see �Fundamentals of Clinical Hematology�, Stevens, W.B. Saunders Company, ISBN 0-7216-4177-6).
Milk quality is essential for farmers, diary producers and consumers. Farmer have to deliver milk of a certain quality which is controlled by the so-called Somatic Cell Count (SCC). In milk quality tests somatic cells in the milk are counted to determine infections (clinical mastitis). A limit of 400,000 cells pr. ml. has to be met by the farmers for dairy resale. Change of diet, stress or mastitis lead to higher SCC levels, thus lowering the quality of the milk and consequently lowering the price per unit volume. A cheap cell counter will help farmers and diary producers monitor SCC-levels.
Samples of milk were obtained through the laboratory of the Danish Dairy Board in Ladelund. Milk contains a large number of fat mono-globules of 5-8 μm size, which are evenly distributed in the fresh milk. In order to count the somatic cells the fat has to be removed or distinguished from the somatic cells. In
Counting with the standard Fossomatic (Foss, Hilleroed, Denmark) and impedance method is not directly comparable. The Fossomatic measures optical signals from propidium-iodide fluorescent marked particles, which is a marker for DNA. Where the Fossomatic only counts particles with DNA, the impedance method differentiates the Somatic cells from their size. However, a small overlapping of fat particles and Somatic cells are present and must be compensated. By subtraction of 10% of the fat count from the Somatic cell count the impedance counting will detect occurrences of mastitis just as well as the Fossomatic (see FIG. 6).
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Whatever its explicit message, Occupy Wall Street has made a powerful statement with its very mode of existence.
The use of clean energy sources is growing, but unless those sources
become cheaper and more efficient, they won't put a dent in the rise in
carbon emissions.
The Century's work relies primarily on subscriptions and donations. Thank you for supporting nonprofit journalism.
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In politics, meeting in the middle is often a useful and necessary thing. But it isn't itself an adequate ethical yardstick.
In the U.S., assisted suicide has mostly been a hard sell. But there are some clear steps to take to improve end-of-life care.
Almost a third of Protestant pastors think domestic violence is not a problem in their congregations. They're wrong.
The question isn't how frightening ISIS is. It's what actual threat it poses—and how to contain that threat without causing more harm.
On election day, the Republicans will keep the House, the Democrats may lose the Senate, and 1,000 more immigrants will be deported.
Behind the Ebola epidemic are issues of basic health care. Combating it involves fairly basic public health measures and education.
Air strikes give the illusion of surgical intervention. But they are not unambiguous humanitarian acts.
The people to whom John Howard Yoder was accountable struggled to discipline him—and failed to deal adequately with his victims' pain.
Instead of seeking the ability to deport Central American children faster, Obama should treat this situation as the refugee crisis it is.
The "war on drugs" approach to marijuana has had major costs. But the dawning era of legal marijuana presents its own set of public health problems.
Sacrifice has real moral resonance—but it can also be exploited. In Iraq, past sacrifices don't offer a guide for U.S. policy.
Readers may or may not accept Charles Hefling's reconstruction of the doctrine of original sin. But he continues the tradition of rethinking the faith in light of new knowledge, contexts, and concerns.
A new study finds that Americans say they attend religious services more than they actually do. Is this bad news for churches?
Legislative action may be slow, but a new consensus is emerging: massive incarceration is unsustainable, both morally and financially.
On the international scene, Rwanda has become a synonym for doing nothing in the face of genocide. After 20 years, what's changed?


text:
ESTERO, FL- An 81-year old woman attacked and robbed in her Estero home. Deputies say she didn't give up without a fight.
Martha Caiceda feels her bruises today. Battle wounds from fighting off a woman she says forced her way into her home and stole her husband's money.
"Here and here, my finger, and my arm," Caiceda said. "She tried to bite me, but I defend me."
Caiceda says Deborah Gardner stormed into her home demanding money for cleaning work she had done previously.
"She said, comprende? Comprende? It's in spanish. I don't know why?"
Caiceda says she'd never seen Gardner before and since her husband was sick in bed, she asked her to leave- but gardner refused.
"I don't care! I need my money," Gardner said.
Deputies say gardner then took the wallet out of Caiceda's husband's pants.
"It's not your money, it's the money of my husband," Caiceda explained.
Then the two began to fight.
"I take her by the hair, I had her head in my hand."
She says then Gardner bit her finger.
"It's very small but it hurt! The woman, take the money, push me down. And go away."
A neighbor called 911.
Deputies say Gardner made her get away on her golf cart, they followed her a few streets away and then got her at her house, in the same community. She was arrested.
Despite her tough actions, Caiceda says she was scared and is now relieved.


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716 results for "nhl lockout"
Kings-Ducks Preview
Kings-Ducks PreviewPreview | May 02, 2014
NHL - NHL, NHLPA have come a long way in a year since the lockout...Scott Burnside
The NHL and the NHLPA have come a long way since the lockout ended, writes Scott Burnside.Story | Conversation | January 17, 2014
NHL - Growing pains necessary for Nail YakupovCraig Custance
ESPN Insider Craig Custance explores the growing process of Edmonton Oilers forward Nail Yakupov, and what we can expect out of him in the near future. He also digs deep to find out the truth about the recent KHL rumors.Blog | Conversation | October 15, 2013
NHL - Hockey Prospectus projections for 2013-14 top goal scorersAdam Gretz
Adam Gretz of Hockey Prospectus projects the NHL's top goal scorers for the 2013-14 season. Tampa Bay's Steven Stamkos tops the list.Story | Conversation | September 24, 2013
NHL - Top 100 prospects for 2013-14Corey Pronman
Corey Pronman identifies his Top 100 NHL prospects for 2013-14, including Jonathan Drouin, Nathan MacKinnon and Seth Jones.Story | Conversation | September 09, 2013
A skeptic's 2012-13 review: the lockoutPaul Grant
After years of following and covering the NHL, some people gain a certain skepticism when it comes to the way the league operates. However, most journalists are able to shake free that skepticism and move on. Not me. Thus, a skeptic's first in ...Blog | July 15, 2013
Despite lockout, NHL not experiencing fallout
Whatever damage was caused by the latest long NHL lockout, the league isn't feeling the fallout as it gets set to start this year's playoffs.Story | Conversation | April 26, 2013
NHL - Ways to improve the Stanley Cup PlayoffsCraig Custance
Craig Custance dives into his mailbag as readers offer up some potential improvements for the Stanley Cup Playoffs.Blog | Conversation | April 19, 2013
Winter Classic between Toronto Maple Leafs-Detroit Red Wings set ...
The Winter Classic between the Toronto Maple Leafs and the Detroit Red Wings at Michigan Stadium, canceled this season due to the NHL lockout, has been rescheduled for Jan. 1 next season.Story | Conversation | April 07, 2013
Time for a little escrow talk (just a little)Pierre LeBrun
Who feels like a little escrow talk? Zzzzzz. Nevertheless, I forge onward. The NHL and NHL Players' Association agreed this week to withhold 10 percent of each player's paycheck this season and put it into the escrow fund. If players' com...Blog | January 25, 2013
Is the end of the NHL lockout good for hockey? - ESPN The Magazin...Chris Jones
Sure, it's great that the NHL is back. But longtime fan Chris Jones wonders if the lockout was this league's last chance to correct the broken, bloated monster it's become.Story | Conversation | January 24, 2013
Thomas Vanek scores two goals as Sabres rally past Flyers
Thomas Vanek scored twice and added three assists in leading the Buffalo Sabres to a 5-2 season-opening win over the Philadelphia Flyers on Sunday afternoon.Recap | Box Score | Conversation | January 20, 2013
Teemu Selanne leads charge as Ducks knock out Cory Schneider, Can...
Teemu Selanne had two goals and two assists Saturday to help make Cory Schneider's first NHL season-opening start a short one and the Anaheim Ducks thumped the Vancouver Canucks 7-3.Recap | Box Score | Conversation | January 19, 2013
Los Angeles Kings finally raise banner, honor Sandy Hook family
The Los Angeles Kings raised their first Stanley Cup championship banner Saturday in a joyous ceremony delayed more than three months by the NHL lockout.Story | Conversation | January 19, 2013
Avalanche-Wild Preview
Avalanche-Wild PreviewPreview | January 18, 2013
Penguins-Flyers Preview
Penguins-Flyers PreviewPreview | January 18, 2013


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By Fred Gonzalez
The goods: Key Largo has its standard restaurant fare – usually a local dive that serves great seafood and has a terrific view. The Big Chill, located at mile marker 104 on the bay side, hopes to change that trend. Part upscale bistro, part sports bar, part outdoor lounge, The Big Chill has something for everyone.
When you walk in you're met by a mammoth-sized bar that cries for you to order a martini or rum drink. A turn to your right leaves you looking at the raw bar, which features fresh shrimp and oysters every night. Directly behind the raw bar is one of three dining areas, each perfect for a reception or banquet, and each featuring its own unique décor. As David Tool, one of the principal owners of the restaurant said, "No two rooms are alike. The floors change, the furniture changes, the colors change. You won't see any fish nets hanging here. Nothing that looks like Key Largo." To the right of the raw bar is the sports bar, an upgrade from most in Key Largo, with a pair of pool tables, 30 42-inch HD screens and plenty to choose from on tap.
Outside is just as spectacular, with a sizeable dining deck overlooking an unobstructed view of the bay (perfect for watching sunsets). Just as impressive is the massive tiki bar, with four "slushee" drink machines (perfect for making a pina colada). There's a lounge area adjacent to the swimming pool and surrounded by seven tiki huts that can be reserved ($250) for VIP service on a weekend afternoon. Each tiki hut comes with its own seating and television screens. Also outside is a stage for live bands, which Tool said will be either Southern rock or blues on most nights.
The grub: The Big Chill will be known for its large, tasty steaks. Its menu is meat-heavy, with everything from filets to prime rib. The restaurant boasts one of the largest meat coolers in the Keys, and Tool is convinced his steaks will be among the most savory in the area. Several sauces are available, including melted bleu cheese crumbles that we chose for our prime rib. The restaurant will also feature happy hour daily from 4-6 p.m. with $1 off all beers and house wines and two-for-one well drinks. Signature drinks are $8.25, our favorite being the Mango Mojito.
The Verdict: The Big Chill will be a solid choice for travelers to the Keys, the options around the swimming pool will be enticing to a youthful crowd and the location is perfect for a road trip from Miami (approximage time 45 minutes). However, the success of this restaurant may depend on the support of the locals and the quality of the staff's service.
The Big Chill, MM 104 Bayside, Key Largo. Open Sun.-Thurs. 11 a.m.-11 p.m.; Fri.-Sat. 11:30 a.m.–midnight.
The Raw Bar at the Big Chill.
[1] http://www.miami.com/restaurants/first-look
[2] http://www.miami.com/the-big-chill-1
[3] http://www.miami.com/1826-restaurant-names-new-chef-article
[4] http://www.miami.com/restaurants/nibbles-and-bits
[5] http://www.miami.com/five-great-miami-cooking-classes-article
[6] http://www.miami.com/list-verde-s-nicolay-adinaguev-article
[7] http://www.miami.com/stripsteak-fontainebleau-latest-michael-mina039s-south-florida-mini-empire-article
[8] http://www.miami.com/tamarina-puts-coastal-italian-flavors-downtown-miami-article
[9] http://www.miami.com/ceviche-105-fresh-start-south-beach-article
[10] http://www.miami.com/modern-italian-bistro-moye-opens-brickell-article
[11] http://www.miami.com/babylon-opens-gateway-turkish-cuisine-south-beach-article
[12] http://www.miami.com/ironside-pizza-slinging-neapolitan-pies-miami-article
[13] http://www.miami.com/seven-dials-gives-classic-british-pub-fare-miami-makeover-article
[14] http://www.miami.com/morimoto-rolls-south-beach-style-article
[15] http://www.miami.com/finka-fuses-cuban-korean-and-peruvian-food-west-miami-article
[16] http://www.miami.com/vintro-kitchen-emphasizes-seafood-miami-beach-boutique-hotel-article
[17] http://www.twitter.com/miamicom


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So you want to develop for Windows 8.1? Visual Studio 2013 is HERE
Hot on the heels of Windows 8.1, Microsoft has released the latest versions of its flagship Visual Studio and Team Foundation Server developer tools, albeit only to MSDN subscribers for the time being. Naturally, Visual Studio 2013 offers support for developing Windows Store apps for Windows 8.1, which officially became …
It's almost as if Microsoft doesn't want developers
It's hard to fathom why Microsoft sets the bar so high to software development on Windows. Between killing third-party tools and raising the price (and complexity!) of their own bloated offering, I get the feeling they're trying to lock out everyone but the developers who *have* to develop for Windows and those who see a bigger reward for Windows programs.
Just today I had to quickly write a simple GUI to control some weird USB devices we received. Although I have a VS licence, it was quicker and easier to install Linux in a VM and use Qt and libusb to develop an application. Total time: about 4 hours. Why must every bit of code in Windows turn into a huge, fscking development project? Even OSX is easy by comparison.
Re: It's almost as if Microsoft doesn't want developers
Is it really that expensive though? Not for a full-time professional. I work with arty folk and they have to buy Maya/3DSMax which are much more expensive last time I checked, and then there's Photoshop. FOSS alternatives exist (Blender, Gimp) but people still believe the paid for products are worth the money and personally I think the same is true for VS.
You also have to remember that the VS Express tools are quite capable for serious use, many companies use them for creating commercial products and they are completely free. You can develop in VS without paying anything more than your Windows licenses if you use VS-Express and SVN/Git/etc.
Re: It's almost as if Microsoft doesn't want developers
While no lover of MS, I agree with JDX. Compared to many high end development tools MS ones are good value and the free versions offer a lot of the functionality.
The tools are also very high quality. I often use the Visual C++ to develop small code snippets before porting it to the OS of choice because the debugger is so good compared to say GDB
Re: It's almost as if Microsoft doesn't want developers
@Quxy - eh? Where did you get this from?
When I started playing around with XNA, all the tools were free to download, and I went from nothing to "proto-game" (graphics moving on screen, some user controlled, with basic collisions) within a couple of hours. Since then, I've gone on to develop several games, all with very minimal cost.
In fact, my single biggest outlay to date for my game development - both in terms of time and money - is Android, specifically the licence for MonoDroid, and difficulties installing Monogame and getting it working.
Re: It's almost as if Microsoft doesn't want developers
We're not talking about XNA and quick games, though, we're talking about the tools to develop serious applications - C++ compilers and so forth. Gcc/Clang/Xcode and so forth on other platforms are free.
Also, a quick Google seems to indicate XNA is no longer being developed by Microsoft...
Re: It's almost as if Microsoft doesn't want developers
Free? Who said anything about that? As I said, I *own* the Microsoft development tools needed to do the job.
It's just that trying to use VS to quickly write a simple USB driver and device GUI is like using a city bus to drive to the corner store. It's possible, but a bicycle is quicker and easier.
It's still as ugly as sin
So I'll stick with 2010 until they drop the TIFKAM bonkersness.
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UPDATED Tuesday, July 1, 2014 --- 11:25 a.m.
COLFAX, Wis. (AP) -- A weak tornado that struck last week caused more than $1 million in damage to Colfax schools and the district's athletic complex.
The Chippewa Herald (http://bit.ly/1oktt3o ) reports the tornado touched down briefly twice Friday before reaching the building housing the elementary, middle and high schools.
Besides causing significant roof damage to the building, the tornado also took out two storage sheds in the athletic complex as well as damaging fences, dugouts, bleachers, grounds and trees.
A couple of big vent covers were ripped off the school roof, leaving holes for rain to pour into the building. Some carpeting and windows will need to be replaced.
School Superintendent William Yingst says the goal is to repair all the damage before the start of the football season and the new school year.
Copyright 2014: Associated Press
UPDATED Saturday, June 27, 2014 --- 7:40 p.m.
From our news partner WEAU in Eau Claire
The National Weather Service has amended it's original results on the tornado that went through Colfax.
It now says the tornado was a weak EF1 category and traveled at 85 to 90 miles per hour.
It was around 100 yards wide at its widest. It says the tornado lasted about five minutes, traveling about 2.5 mph.
The twister went through Colfax around 3:15 Friday afternoon.
UPDATED Friday, June 27, 2014 -- 6:48 p.m.
COLFAX, Wis. (AP) -- A weak tornado has struck the western Wisconsin village of Colfax, downing trees, tearing roofs off homes and knocking out power.
Police Chief Bill Anderson tells The Associated Press the brief tornado struck without warning "right on main street" around 3:15 p.m. Friday.
Anderson says the tornado damaged a gas station and tore roofs off some houses, but only one family was displaced. No injuries are reported.
The storm also twisted goalposts at the high school football field and ripped venting off the school roof.
National Weather Service technician Ross Carlyon tells the Leader-Telegram it was a "very fast-moving thunderstorm" that briefly dropped a weak tornado.
Colfax is a village of about 1,100 about 75 miles east of Minneapolis. In June of 1958, a deadly tornado nearly obliterated Colfax.
Copyright 2014: Associated Press
Posted Friday, June 27, 2014 --- 6:02 p.m.
COLFAX, Wis. (AP) -- The National Weather Service says a weak tornado has struck the western Wisconsin town of Colfax, but damage reports are minor.
Law enforcement tells the weather service the tornado damaged a gas station and knocked down trees Friday afternoon. No injuries are reported.
Weather service technician Ross Carlyon tells the Leader-Telegram (http://bit.ly/1lVVTAF ) the tornado hit without warning. Carlyon says it was a "very fast-moving thunderstorm" that briefly dropped a weak tornado.
The tornado hit around 3:15 p.m.
Colfax is a town of about 1,100 about 75 miles east of Minneapolis.
Copyright 2014: Associated Press


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Minnesota > Moorhead Yellow Pages > Industrial Goods and Services > Industrial Materials: Metals Moorhead
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November 22, 2014
Several people were hospitalized, including one taken by medical helicopter, following a two-vehicle accident in Omaha Friday morning.
A car and SUV collided at South 204th Street and George B. Lake Parkway just after 9 a.m. The traffic lights at that intersection have not yet been activated and are covered in plastic.
No word on the extent of the injuries.
Click Here to access the online Public Inspection File
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We asked former Ticket board op and master nicknamer Michael "Grubes" Gruber to autopsy the Rangers so we can throw their cadaver into our winter fire and focus on football. Here's his hot-but-not-tremendously-hot sports take: This was a weird season for your Texas Rangers. For all intents and purp ... More >>
Photos by Sam MertenNelson Cruz giving me a free ticket to the gun show last year.As the trade deadline approached in July 2006, it seemed inevitable that Jon Daniels would make a bold move to bolster a mediocre offense in his first season as general manager. He targeted the best hitter on the mar ... More >>
As we mentioned yesterday, Jon Daniels is entering his fourth year as Texas Rangers general manager. He started off with a bang in year one, trading Alfonso Soriano in his first move and dealing Chris Young and Adrian Gonzalez less than two weeks later. Unfortunately, he received very little in retu ... More >>
Okay, not really. But we can dream, right? Because if Pacman Jones did get his own show with The Ticket's informative and entertaining Bob Sturm and Dan McDowell, the corrupt Cowboy returning to practice at Valley Ranch today would mysteriously fall on hard times and ... More >>
Seven-year-old Caroline Turns, who was recently diagnosed with a rare form of pediatric cancer Not sure what's more mind-blowing to Kelly Howell these days. the fact that a precious 7-year-old girl can get pancreatic cancer. Getting up at 5 a.m. to run 13 miles. Or the kindness of total strangers. ... More >>
Another season, another last-place finish, another shrug, another yawn. In the wake of an obscure 75-87 disappointment that has already camouflaged itself alongside the other 22 losing seasons in franchise history, you'd like to hear something drastic or at least different from the Texas Rangers. ... More >>
Is baseball's youngest general manager man enough to rebuild the Rangers?
Can the Rangers' oldest rookie leave his diva days behind and deliver?
New Rangers manager injects hope and, yep, even optimism in Arlington
www.bobanddan.com If you see Dan McDowell, left, and Bob Sturm coming, run the other way. Drew Bledsoe didn't; look what happened to him. Fonzie, yer next. According to popularity polls, the Fort Worth Star-Telegram and totally illogical thinking, Tony Romo will today be officially named the Dallas ... More >>
Whenever we see a picture of Chris Young in a Padres uniform, we cry just a little. Then we drink and cry some more. Then we drunk-dial Adam Eaton. Since firing Texas Rangers manager Buck Showalter a week ago, general manager Jon Daniels has been quiet about a replacement. Hitting coach Rudy Jaramil ... More >>
Another Rangers season ends in a familiar Hunt for Dead October
It's an annual rite of spring. On April 15--at precisely 11:59 p.m., if you're like me--we do our taxes and write off the Texas Rangers. This year seemed no different. Before the season even started, prized pitcher Adam Eaton injured his finger. New leadoff man Brad Wilkerson started striking out mo ... More >>
The Texas Rangers have played 13 games. It's the equivalent of the Dallas Cowboys having just gone into overtime in their season opener. So should those of us who care about the local baseball team be as miserable after the 5-8 start, especially before the just-concluded trip to Oakland, as the loca ... More >>
Is this the year the Texas Rangers finally...? Nah.
A young dog teaches the Rangers to forget old tricks


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Watercress and Fennel Salad with Blood Orange and Thyme Vinaigrette
Posted: Updated:
Watercress and Fennel Salad with Blood Orange and Thyme Vinaigrette
Gentl & Hyers
Provided by: Taste Editors
total prep
The combination of bitter greens, soothing fennel and tart blood orange makes this a super-refreshing and beautiful addition to nearly any spring menu.For a completely different taste, substitute fresh mint leaves for the thyme, or use both!
Recipe from Clean Start by Terry Walters/Sterling Epicure, 2011.
Recipe from Clean Start by Terry Walters/Sterling Epicure, 2011.
- 2 blood oranges
- 2 bunches watercress, trimmed
- 1/2 fennel bulb, cored and thinly sliced
- 1/4 cup lemon juice
- 1/4 teaspoon lemon zest
- 1/2 cup extra virgin olive oil
- 1 tablespoon minced shallot
- 4 teaspoons maple syrup
- 1 tablespoon fresh thyme leaves, chopped
- Sea salt and freshly ground pepper
- To prepare oranges, cut off both ends, and cut between the peel and the orange to remove peel and pith completely. Cut oranges crosswise into 1⁄4-inch slices, place in bowl and set aside.
- Arrange watercress on individual plates or on a large platter and top with sliced fennel. In small bowl, whisk together lemon juice, lemon zest, olive oil, shallot, maple syrup and thyme leaves. Season to taste with salt and pepper. Remove oranges from bowl and arrange on salad. Pour remaining juice from oranges into the vinaigrette bowl. Whisk to combine, drizzle evenly over salad and serve.


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the characteristics, syndrome, or diagnosis of a nigga-- tardiness, stingyness, laziness, mooching, greediness, irresponsiblity, undependability, and falling asleep after eating. Similar to an illness.
The rap concert started late, cause Lil'Wayne was suffering from a bad case of nigga-i-tis
by treece December 22, 2005
The syndrome that may afflict certain people after they smoke weed. Sometimes, after smoking, one will feel tired and lazy, only wanting to sit around and do nothing. This phenomenon has been associated with the stereotypical black person, or nigga. May also be referred to in conversation as
"the itis"
"the itis"
Guy 1: "Dude, I'm so stoned right now. I can't even get up off my ass."
Guy 2 (to friend): "Look at him bro- niggaitis strikes again!"
Guy 2 (to friend): "Look at him bro- niggaitis strikes again!"
by stoner44 August 03, 2009


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Dolby Laboratories, Inc. (NYSE:DLB)
Dolby Laboratories develops and delivers innovative products and technologies that make the entertainment experience more realistic and immersive.
When the market is crashing and you are reporting earnings, then sometimes you get stabbed a few extra times with the skewer. Dolby Laboratories, Inc followed the markets the first part of the week, down about 10% going into earnings. When you factor in the other 18% on Friday after the earnings report and compare it to the peak in January, you're down over 50%. Since Dolby Labs has been a Motley Fool Stock Advisor pick three times sine it debuted in 2005 I guess I shouldn't be surprised to see the Motley Fool bashing that has started after the plunge. It seems some people just love it when they are eventually proven right and someone else is wrong.
In this case, it certainly pays to have a diversified portfolio as earnings data this cycle has resulted in a wave of very measurable price changes. I'm certain I can find 50 equities that have either gone up more than 10% or down more than 10% on earnings day this cycle. This certainly goes to show how reliable analysts and market pricing is.
That Dolby reported in a market crushing two day period definitely increased the number of lashes the market dispensed. It also meant that the interpretation of the reports was done much more hastily as focus was on the blood letting going on in the rest of the market. Dolby reported near in-line with a flat EPS of $0.55. Flat lately is apparently bad. The hasty part, however, may have been the interpretation that so far, Dolby Labs is not an automatic licensing shoe-in for Windows 8. While Window 7 revenue has been about 25% of Dolby's revenue and it's loss might warrant some lashes. The reality, however, is that:
1. This will not affect Dolby until at least 2013.
2. Dolby, if not included can still license it's products to the PC makers, such as Dell, HP etc.
3. Dolby's PC sales are a fraction of it's total placement which is growing.
When you measure Dolby's cash flow and earnings they have shown solid growth EVEN during the 2008-2009 recession. (Over 15% increase between 2009 and 2010) and 31% profit margins the last year. (The flat EPS was partially due to accounting changes between 2010 and 2011). Factor in the $7 per share cash, no debt, the solid cash flow, and the stock buybacks, (just renewed) and you have a company that is about as double dipped recession proof as you are going to find.
Now back, briefly, to the criticism of the Stock Advisor, (the flagship MF paid service) recommendations.
1. I think the sell off was overdone because their was no time to interpret the Microsoft Relationship.
2. The earnings came on a down day, accelerating the drop.
3. If you had bought Dolby when in 2005 you'd still, (after a 50% seven month drop), be kicking the S&P by about 30%. Certainly not as much fun as the 150% you might have been in January, but the ride is not over.
4. If you were diversified you probably wouldn't have felt the motion sickness quite as much.
Fortunately, for me, I can be a little upbeat as I didn't own Dolby before the selloff. I have this phobia about buying something that is doing well. I'm not sure if it's caused by:
1. The fear that I've already missed out and there isn't as much gain left for me.
2. I'm jealous of those who did invest.
3. I have this subconscious desire to hurt myself.
So what does that mean now? I don't know, if I did I'd be unsuccessfully running a hedge fund somewhere, (see 3 above). My meager trading/investing experience, especially in the crashes of Oct '08 to March '09 is that MANY of the equities that got hammered in the earlier rounds of the sell off were actually above their 2008 lows when the bottom settled in 2009. Those equities that got overly punished built their base and rode the waves. Name brand company's with strong earnings were quick to recover and exceed their previous levels.
Dolby was $42.61 Sept 15th 2008, $25.56 March 9, 2008, then $67 Jan 1st, 2011. It not only bounced back, it excelled, because all through the recession it continued to grow.
No anyone who looks at a company's past performance and things it's a reflection of the future isn't going to last very long in the investment world. Many company's don't innovate or get squeezed by newer technology or competition. Looking at past numbers are useful, but shouldn't be the basis of an investment decision.
IN the case of Dolby, however, there is nothing in the history that tells us that they are subject to a double dip recession anymore than anyone else. In actuality, they have proven they can ride through it. Dolby is an innovator and even in a recession there are some things consumers will stretch for. I thought in the last recession that auto makers and auto parts makers were in trouble. Was I wrong! It seems that when people lose their jobs they will give up their houses, but how many gave up thier cars or thier stereos? Look around...how many cars are in the drive way at Mom/Dad's where the kids moved in? How many in front of the three decker apartments where those foreclosed on moved into.
So, for me, it's a chance to get a piece of an innovator, solid cash producer, at a bargain price. If things don't go optimal for Windows 8 in the next two years I'll see if Dolby makes up for some of it with the PC makers. If not, I still have a cash machine buying back its shares. I fully believe that market selloffs have no mentality in choosing their victims; the market in these cases is extremely inefficient. The more I can leverage the inefficiencies the less risk I will have during further panic. If Dolby drops further in more market selling or more mis-interpretation of the reports then who am I going to blame?? No one, not even myself. I'm going to continue researching, evaluating and investing Dolby and other equities. I'll diversify. I certainly won't blame the Gardner brothers if I buy Dolby after doing my own DD, I don't see them forcing me to be so Foolish.
Just how trashed does a stock price have to get before The Fool sells?
I think Mr. Dolby is paying them off to keep them on the buy list.
Also, if you want to be sitting on huge losses in another Fool favorite,
keep holding Netflix. The Fool would be wise to dump it today.
I don't know, you can try both of those questions on the SA boards where both are favorites. It doesn't appear that too many folks are "happy" about multiple favorites falling so soon after each other, but both are still up considerably over their first recommended prices. I think Netflix is in worse shape, but if you bought either at the peak, it's been a tough month.
I don't believe either are paying anyone off. They both had models that fit the SA principles. SA is particularly fond of the subscription model like Netflix and licensing such as Dolby. When they work, the margins are usually good and addional sales are usually at higher margins. Good luck.
Just for fun I'll guess that it'll bottom at $18.91. Seems like at $27 bucks today no one thinks the company will ever grow. I think the bad news is priced in. When they start growing again or offer great guidance, the PE ratio will shoot right up again. Best of luck shareholders. If anyone wants to sell me their shares, I'll offer 20 bucks each. Please overreact.
I sold Dolby in May after a 25% loss. Thought at the time it would shoot straight back up to 65 because I sold it at 50. Took it completely off my watch list at that time and lost track of it until I read some of your comments. Was real glad I sold after seeing the reaction to 2Q earnings on Aug 4th. Perhaps its time to get back in! It certainly would be a contrarian play at this point.


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Welcome to the ThingWorx site. We're excited to have our message and company out in the public domain and are confident that ThingWorx will become the center of gravity in the new Internet of Things.
IBM is right in linking the success of the Internet of Things to Man's ability to create a smarter, more sustainable planet. History has shown that great advances, such as the transition from the industrial age to the information age, can't happen without the introduction of a disruptive technology.
ThingWorx is it! Its revolutionary design will provide the disruptive change required to catalyze the Internet of Things and lead to a more efficient and innovative world.
You'll be hearing a great deal from us over the next months so stay in touch, follow us on Twitter, and join us in our goal to remake the planet, one thing connected business process at a time!
Russ Fadel


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Series 7 Filming: 60s Dalek on Set
It appears the BBC really did mean "all" Daleks will be returning when they announced the news on Friday.
Today, Steven Moffat tweeted this set pic of Matt Smith and Karen Gillan posing with a dusty Dalek model from 1967's The Evil of the Daleks!
Moffat said: "Matt Smith likes the sixties Dalek best."
Step back in time...
COMMENT GUIDELINESPlease be civil and keep article comments relevant and on topic. Flag and report any offensive/trolling behavior, or contact us with details.
Please do not post SPOILERS including anything from leaked episodes! Your account could be banned. For complete details on our comment policy please read.


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Tradition requires a year-end listicle. We could just tick off the news events to which rightbloggers reacted ridiculously over the past 52 weeks -- from gay marriage victories to Rand Paul's filibuster to the Boston Marathon bombing to the George Zimmerman trial to Obamacare. But everyone else i ... More >>
Yesterday's stars at today's prices
Stuck in the asylum with the good kind of insane
Film probes the mutual codependency between fan and fame
Unintentionally playing the roles of Rupert Pupkin and Masha in The King of Comedy
One Guy Tries Harder for Brooklyn Borough President


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Ratings based on 5 Verified Reviews
They paid and stayed. We double-checked.
3.6 Room cleanliness
3.6 Service & staff
3.4 Room comfort
3.8 Hotel condition
They paid and stayed.
We double-checked.
We double-checked.
Recommendedby A verified traveler
Posted May 20, 2014 on Hotels.com
for Business Travelersby A verified traveler
good hotel, value for moneyPosted Jul 23, 2013 on Hotels.com
Nice hotel, should have a better overall score.
for Couplesby A verified traveler
Good pricePosted Oct 1, 2012 on Hotels.com
Was ok, the only thing is that they hav to put a door between the kitchen and the bed space, when you cooke everything smell.
Recommendedby A verified travelerfrom Camberley
Get a code before staying here.Posted Apr 4, 2011 on Hotels.com
If you are looking for basic overnight accomadation this is GVFM. The appartment is close to a supermarket and fast food outlets.The issues arise if you arrive at the weekend, you need a code to get your key from the safety deposit box. No reception at the weekend means you are on your own. 30 frustrating minutes later we got the code. Still a good stopover on the way to the Alps.
Recommendedby A verified travelerfrom London
Excellent valuePosted Apr 24, 2009 on Hotels.com
For an essentially unattended hotel, the service was excellent. The receptionist was exceptionally helpful, not only with check-in, but directions, maps and internet access


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Web Service DeveloperVacancy posted: Saturday, 19 January 2013
Salary: £25-£30K Plus Excellent Benefits
Location: Glasgow
Please note: This vacancy is no longer advertised.
You might find these similar jobs of interest:
- Test Analyst, Carlyle Associates, Edinburgh
- QA Engineer - newly formed team , Cathcart Associates, Glasgow
- Digital Software Engineer, FPSG Connect, Edinburgh
- QA Engineer, Eden Scott, Edinburgh
- Software Automation Tester, HAYS, Glasgow
Web Service Developer
Based in Glasgow
Salary £25k-£30k Plus Excellent Benefits
Our client a market leader in Financial Services have an exciting opportunity for a Web Service Developer to join their highly successful organisation.
Working with the Technical Architect to design, build, test and implement solutions to integrate business applications using web services and other relevant technologies. Identifying the correct technologies to be utilised on various projects. Working with the Technical Architect to design solutions and work through the full system life cycle.
Additional Tasks:
- Production of solutions to required standards.
- In conjunction with other members of IT, help with development of appropriate development standards.
- Keep up with trends in technology through use of the internet, IT magazines, attending workshops and conferences etc.
Key Skills and Experience Required:
- Degree level (or equivalent) in computing or related subject.
- 1 to 2 years Java experience.
- Desirable: XML/XSLT, Web Services.
- Beneficial: Linux, JMS, SOA, ESB, SQL.
- Ability to quickly pick up new technologies and techniques.
- Effective communication and interpersonal skills.
- Practical experience of the full development lifecycle.
- Strong Relationship building skills.
- Ability to develop alternative solutions to issues and drive through improvements
- Proactive approach
- Clear & concise report and documentation writing skills
If you are confident you have the relevant skills and experience required for this role then please forward your CV in the 1st instance to email@example.com
About Suzanne Paterson Consulting
Suzanne Paterson Consulting offers bespoke recruitment solutions across a range of Commercial Sectors, including Call Centre, HR and Executive Search. Having 12 years recruitment industry experience,we provide a quality service to both clients and candidates alike.We aim to tailor recruitment solutions for each client to ensure results are achieved efficiently and successfully. As an Independent Consultancy you can be assured of a professional, personal and attentive service at all times.
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Tink's® #69 Doe-In-Rut® Buck Lure 4 oz. bottle
Item # WX2-110126
- Google+
This item cannot ship via any of our Next Day or Second Day Express delivery methods.
You must be 18 years of age to purchase this item.
Shipping restrictions apply to this product under some jurisdictions. Known age, state and local shipping restrictions are applied at checkout and may result in changes to an order. PLEASE NOTE: We do not accept returns of Ammunition for safety reasons. NEVER use any Ammunition unless it is specifically designed for your firearm.
This item cannot ship via any of our Next Day or Second Day express delivery methods.
This item cannot ship to Canada.
This item cannot ship internationally.
- Details & Specs
Tink's ® #69 Doe-In-Rut ® Buck Lure.
The original Doe-In-Rut ® Buck Lure is a blend of 100% natural aged doe estrous urine which is specially formulated to keep product at the peak level of performance. Special odor comes from Tink's secret, quality controlled formula in an easy-to-use squeeze bottle.
Try America's best-selling Buck Lure for yourself! Order ONLINE Today!
- Reviews
Tink's #69 Doe-In-Rut Buck Lure 4 oz. bottle is rated out of 5 by 2.Rated 5 out of 5 by KIRK & LORI from i switched to tinks about a year ago was using code blue. my brother turned me onto tinks great hunts but will still use coe blue when neededDate published: 2013-10-12Rated 5 out of 5 by DAVID from Tink's has been around since i was a kid, I've been hunting with #69 for 38 yrs. , gotta be a reason for that.Date published: 2011-09-28
<!--BVSEO|dz:Main_Site-en_US|sdk:v188.8.131.52-n|msg:Error: Request errored out - The remote server returned an error: (403) Forbidden. -->
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text:
The extracts on this page are taken from selected national newspapers. The views expressed here do not necessarily reflect the views of Manchester United.
Ian Herbert writes in The Independent...
Sir Alex Ferguson appears to be convinced he can overcome Arsène Wenger's resistance to losing Samir Nasri and capitalise on the French midfielder's contractual impasse with a speculative £20m offer. As far as Wenger is concerned Nasri is not for sale and ostensibly his price is the same £40m that Arsenal would want for Cesc Fabregas – because the Arsenal manager wants him out of Ferguson's reach. But United, whom The Independent revealed three weeks ago have held initial discussions through intermediaries of Nasri, have yet to test how Arsenal will respond to a £20m offer for a player who is out of contract next summer and thus worth probably half that sum. United's only option now is to make an offer.
Many papers carry quotes from David de Gea's MUTV interview, which you can watch on ManUtd.com. The Daily Mail also report: "After speaking to MUTV, De Gea flew back to Madrid for a press conference to say goodbye to Atletico. He was overcome by emotion as he tried to explain his reasons for leaving. The Spain Under 21 goalkeeper broke down in floods of tears as he bade farewell to his boyhood club, with whom he won the Europa League in 2010, beating Fulham in the final."
Elsewhere, The Sun claim Sunderland are ready to step up their pursuit for Reds trio John O'Shea, Darron Gibson and Wes Brown.
More: The Gossip | Nani committed to Reds | Macheda vows to learn


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Tears For Fears
'Glee' Gets A Makeover With Help From Sarah Jessica Parker
This week's episode features music from Tears For Fears, Hole and Sheryl Crow.
Flashback Friday: Tears For Fears – Everybody Wants To Rule The World
Remember this song from 1985?
Friday 80′s lunch preview
Some of the best songs in pop music came from the 80′s. Everyday, we play the 80′s Lunch at noon. Click 'more' for a preview of tomorrow's 80′s Lunch.


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Dream Dictionary
Welcome to Dream Dictionary!
You may expect a rash of petty annoyances over money matters if your dream involved sliding down. Going up, in any manner, signifies hard effort before reaching your goal. If the balustrade was broken or incomplete, there could be serious obstacles to some current plan.
Dream Decoding 101
Take our crash course in dream interpretation!
Bedside Notes
Print this notepaper to jot down dream details before they fade away.
Need help?
Step-by-step instructions for using Dream Dictionary


text:
Table 2
Population-based studies of pre-stroke and post-stroke dementia
L=longitudinal. X=cross-sectional. V=volunteer cohort. RPS=retrospective patient study. PPS=prospective patient study. H=haemorrhagic stroke. IS=ischaemic stroke. Y=yes. N=no. DSM=diagnostic and statistical manual of mental disorders. MMSE=mini-mental state examination. NEMESIS=North East Melbourne Stroke Incidence Study. ..=data not available.
* Mean of total volunteer sample.
† Median of total volunteer sample.


text:
Featured news
Virus writers moving faster with attacks
Thursday, 18 August 2005, 6:33 AM EST
U.S. media companies and other corporations hit by a wave of computer viruses this week said business was back to normal today, but analysts warned the attacks showed that hackers have gained a dangerous advantage in speed in the battle over network security.
By Spencer Swartz at Computerworld.
[ Read more ]


text: Munson Mania Arizona!! View full-size image < previous next > Is this photo a winner? "Twelve of us came from New York to Arizona to visit Glenn - son, brother, uncle - depending upon who you were! Had a blast and can't wait to go back again!" - Munson Mania Arizona 2012 (Oct 19, 2012) Share this photo on: Twitter Facebook Pinterest Similar Photos: arizona | event reunion | event vacation | family | family reunion | fun | happy | pool | yellow Feeling inspired? Visit the CustomInk Design Lab to start your own group order. Design Your Own


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David Letterman Apologizes to Wife Regina Lasko, Late Show Staff on Airby Free Britney at . Comments
Taping his first episode of The Late Show with David Letterman yesterday since his stunning admission last week that he'd had sex with female employees, the host apologized to his wife, Regina Lasko, and his entire staff on the air.
It may not instantly fix his reputation, but it was heartfelt.
Of Regina, he said "She has been horribly hurt by my behavior, and when something happens like that, if you hurt a person and it's your responsibility, you try to fix it."
"At that point, there's only two things that can happen.
"Either you're going to make progress and get it fixed, or you're going to fall short and perhaps not get it fixed, so let me tell you folks, I got my work cut out for me."
The late night host married Regina Lasko, with whom he has 5-year-old son Harry, on March 19 of this year in Choteau, Montana, after more than 20 years of dating.
Presumably, although details remain unclear, he was dating Lasko at the time of his office relationships with Stephanie Birkitt, Holly Hester and possible others.
Letterman also took the time to apologize to the entire staff and Worldwide Pants production company, saying "I'm terribly sorry I put the staff in that position."
"The staff has been wonderfully supportive to me," he added, thanking them for "once again, putting up with something stupid I've gotten myself involved in."
CBS was quick to post the clip online (having yanked Letterman's admission from last week). Dave ended his apology by apologizing to ... Sarah Palin. Funny.
Dave reiterated that despite his regrettable and inexcusable behavior, he is the victim of an (alleged) crime, and was being blackmailed about these affairs.
Typically, the 62-year-old also tried to find whatever humor there is. In his opening monologue, he began with three shots at himself in about 30 seconds:
- "Didn't the weekend just fly by?"
- "I tell ya, right now I'd give anything to be hiking on the Appalachian trail."
- "You know, I got into the car this morning and the navigation woman wasn't speaking to me."
Follow the jump for a video of David Letterman's apology ...
she shouldnt forgive him.
By keeping Letterman, CBS has become Columbia Broadcasting Sex.
I really feel bad for his wife!
WOW never saw that coming!
now he is sorry....hahhahaha
It really shoudn't be this big of a deal, he slept with women on his show before he was married however he was still with Regina, so my suggestion would be to stop saying your sorry to her on national TV I think you have embaressed her enough work on your "im sorrys" at home without a camra.
This poor guy. He had sex with a woman when he wasn't exclusive with anyone else. That woman happened to work for him. Big freakin deal. He shouldn't have to apologize to anyone, let alone during his own show. The guy who tried to extort him is a criminal and should be tried as such. HE's the bad guy here, not Letterman.
What is the big deal what late night talk show host or star asn't had an afair. Come up with something better. I do have to say this the girl that had an afair with him must of been hard up!!