Abstract: Methods and devices for evaluating a sample, e.g., a plasma sample, from a subject, for detecting a target red blood cell protein or antibody are disclosed. In one embodiment, optimized antibody screening methods and devices significantly reduce the level of non-specific binding to a surface (e.g., a test surface bound with a red blood cell (rbcm) preparation), thus allowing for more efficient detection and reduced test time. In one embodiment, the optimized antibody screening method includes an immunoglobulin G (IgG) binding moiety that binds selectively and specifically to the plasma IgG present relative to the binding to the lysed rbcm preparation. In another embodiment, an optimized antibody screening method is disclosed whereby non-specific binding caused by lysed red blood cell membrane preparations can be reduced by an agent that specifically cleaves a human IgG in the hinge region.

Abstract: The present invention relates to methods and apparatuses for the detection of positional freedom of particles used in biological, biochemical, physical, biophysical, and chemical analyses. In particular, the present invention relates to methods and apparatuses which can detect and characterize a population of particles/cells based upon their detected mobility. In one embodiment consistent with the invention, detection of certain cells is based on differences detected in populations of cells that bind to a substrate and those that exhibit weaker binding forces. Initially, cells are settled on the substrate, and in the presence of gravitational, natural thermodynamic pressure fluctuations, and other random or applied forces, some of the particles may exhibit translational movement. Particle movement is detected, and measurements are computed, according to the methods and apparatuses of the present invention, to determine the binding of specific analytes.

Abstract: A rotor is provided for use in a centrifuge system configured to spin the rotor for separating complex fluids. The rotor includes a housing configured to be secured by the centrifuge system and several chambers formed in the housing. Each chamber includes a first chamber portion having a port formed therein and a second chamber portion in fluid communication with the port of the first chamber portion. The second chamber portion may be disposed generally below the port of the first chamber portion. Other rotor designs and methods for separating complex fluids are further disclosed.

Abstract: The present invention relates to methods and apparatuses for the detection of positional freedom of particles used in biological, biochemical, physical, biophysical, and chemical analyses. In particular, the present invention relates to methods and apparatuses which can detect and characterize a population of particles/cells based upon their detected mobility. In one embodiment consistent with the invention, detection of certain cells is based on differences detected in populations of cells that bind to a substrate and those that exhibit weaker binding forces. Initially, cells are settled on the substrate, and in the presence of gravitational, natural thermodynamic pressure fluctuations, and other random or applied forces, some of the particles may exhibit translational movement. Particle movement is detected, and measurements are computed, according to the methods and apparatuses of the present invention, to determine the binding of specific analytes.

Abstract: The present invention relates to producing more densely functionalized indicator cells which along, with other components, can be used to detect the presence or absence of antibodies or antigens, by using the A, B, AB and MNS antigens which have a greater degree of expression than the conventionally used D antigen to label the indicator cells with IgG. The present antigen systems have levels of expression that approach one million antigens per cell, which is in great contrast to the conventional D antigen sites of about 10,000-30,000 antigens per cell. This marked increase in the antigen systems level of expression could produce a boost in the kinetics and the magnitude of the binding of the indicator cell to the solid phase, which improves assay performance.

Abstract: The present invention relates to an instrument and a measurement apparatus and methodology that yields a measurement and test methodology that characterizes a population of cells/particles or detects a sub-population of cells/particles based on their detected mobility in a quick and efficient manner.

Abstract: Methods and devices for evaluating a sample from a subject for detecting a target red blood cell protein or antibody are disclosed. In one embodiment, optimized antibody screening methods and devices significantly reduce the level of non-specific binding to a surface (e.g., a test surface bound with a red blood cell preparation), thus allowing for more efficient detection and reduced test time. In other embodiments, the invention provides methods and devices for target capturing that include a substantially planar surface, optionally having an optimized angle, for capture. Alternative solid phase geometries for capture are disclosed. Optimized methods for cell deposition are also disclosed. Thus, optimized methods, devices, kits, assays for evaluating a sample are disclosed.

Abstract: The invention provides a method, apparatus and system for separating cellular components, and can be combined with holographic optical trapping manipulation or other forms of optical tweezing. One exemplary method includes providing a first flow having a plurality of components; providing a second flow; contacting the first flow with the second flow to provide a first separation region; and differentially sedimenting a first cellular component of the plurality of components into the second flow while concurrently maintaining a second cellular component of the plurality of components in the first flow. The second flow having the first cellular component is then differentially removed from the first flow having the second cellular component. Holographic optical traps may also be utilized in conjunction with the various flows to move selected components from one flow to another, as part of or in addition to a separation stage.

Abstract: Methods and apparatus for analyzing surface properties of particles are provided. A method for analyzing the surface properties of the particle includes a associating a first particle with a first capture zone having a specific binding affinity for a first chemical species, applying an optical force to the first particle, sensing a response of the first particle to the optical force, and using the sensed response to determine the presence, absence or quantity of the first chemical species on the first particle surface. This process may be repeated in parallel to test multiple particles. In addition to directly testing the surface properties of the particles, the method can be used in direct, indirect and competitive assays to determine the presence, absence or quantity of free or immobilized analytes. A fluidic cartridge with capture zones having avidities that are tuned for the use of optical forces is provided. A software routine for performing the method is also provided.

Abstract: The invention provides a method, apparatus and system for separating blood and other types of cellular components, and can be combined with holographic optical trapping manipulation or other forms of optical tweezing. One of the exemplary methods includes providing a first flow having a plurality of blood components; providing a second flow; contacting the first flow with the second flow to provide a first separation region; and differentially sedimenting a first blood cellular component of the plurality of blood components into the second flow while concurrently maintaining a second blood cellular component of the plurality of blood components in the first flow. The second flow having the first blood cellular component is then differentially removed from the first flow having the second blood cellular component. Holographic optical traps may also be utilized in conjunction with the various flows to move selected components from one flow to another, as part of or in addition to a separation stage.

Abstract: The present invention utilizes a holographic optical forcing array for dynamic cellular probing and diagnostics. A holographic optical trapping system generates optical forces on objects so that deformations thereof may be quantified. In one embodiment, digital holography is used to generate an interference pattern, and an analysis thereof determines the phase profile which yields a measurement of the objects' shape deformation using only one image. In another embodiment, phase-stepped holography allows the phase profile of an object to be measured using only one image, by using a holographic optical element to make phase-shifted replicas of the beam in space. In another embodiment, the optical forcing array applies optical forces to beads placed on the objects' surface, deforming the objects. The beads' position is determined by applying Mie theory, and analysis thereof yields the three dimensional position of the beads, and a measurement of the deformation displacement on the objects' surface.

Abstract: The present invention relates to a method and apparatus of sorting objects including, providing a sample having wanted objects and unwanted objects; coating a surface of a sample holder with an antibody; placing an eluted sample on the sample holder; binding an antigen in the wanted objects with the antibody on the surface of the sample holder to sort the objects into wanted objects and unwanted objects; separating the wanted objects; and performing PCR-based STR analysis on the wanted objects. In one embodiment, holographic optical trapping is used to further sort the wanted objects. In other embodiments, the wanted objects are sperm and the antibody is a human sperm specific antibody, and the PCR is single cell PCR-based STR analysis. In still other embodiments, the binding is direct or indirect, ligands are used to bind to object-specific organomolecules, and protein A or protein G are used to bind the antibody.

Abstract: The present invention relates to a method and apparatus of sorting objects including, providing a sample having wanted objects and unwanted objects; coating a surface of a sample holder with an antibody; placing an eluted sample on the sample holder; binding an antigen in the wanted objects with the antibody on the surface of the sample holder to sort the objects into wanted objects and unwanted objects; separating the wanted objects; and performing PCR-based STR analysis on the wanted objects. In one embodiment, holographic optical trapping is used to further sort the wanted objects. In other embodiments, the wanted objects are sperm and the antibody is a human sperm specific antibody, and the PCR is single cell PCR-based STR analysis. In still other embodiments, the binding is direct or indirect, ligands are used to bind to object-specific organomolecules, and protein A or protein G are used to bind the antibody.

Abstract: The present invention relates to an apparatus and method of sorting objects and identifying the objects in a forensics sample, including using holographic optical trapping to sort objects from contaminants, and performing (single cell) PCR-based STR analysis on the objects to determine their identification. In addition, the chip used as a support for sorting the objects can also be used for performing single cell PCR-based STR analysis. In another embodiment, a microfluidics chip is used to stream the sample and sort the objects, before single cell PCR-based STR analysis is performed. The chip used for sorting utilizing HOT in the absence or presence of microfluidic streaming and sorting can also be the same as that used for the single cell PCR-based STR analysis.

Abstract: The present invention relates to an apparatus and method of sorting objects and identifying the objects in a forensics sample, including using holographic optical trapping to sort objects from contaminants, and performing (single cell) PCR-based STR analysis on the objects to determine their identification. In addition, the chip used as a support for sorting the objects can also be used for performing single cell PCR-based STR analysis. In another embodiment, a microfluidics chip is used to stream the sample and sort the objects, before single cell PCR-based STR analysis is performed. The chip used for sorting utilizing HOT in the absence or presence of microfluidic streaming and sorting can also be the same as that used for the single cell PCR-based STR analysis.

Abstract: A fluidic device includes an arrangement of channels for introducing a sample containing particles of interest into a processing chamber. The chamber is in fluid communication with collecting channels via low-flow connection channels. Particles in the sample may be observed and diverted from the processing chamber by application of a motive force such as optical trapping into a collection channel. Once in the collection channel, particles can be collected, including by trapping in a porous matrix.

Abstract: An apparatus and method for purifying a forensic sample using an automated extraction and purification system includes optical tweezers; an input channel through which the sample is introduced; a chamber which receives the sample from the input channel; a collection channel through which selected particles of the sample are removed; and an output through which unselected particles of the sample are removed. At least one buffer input channel is provided. The input channel may allow sedimentation of the sample into the chamber by gravity. In another arrangement, the system includes an optical trapping system; a first channel through which the sample is introduced; a second channel through which buffer is introduced; wherein the optical tweezers are used to move selected particles of the sample from the first channel to the second channel. The selected particles may be sperm. The optical tweezers preferably utilize holographic optical trapping.

Abstract: An apparatus and method for purifying a forensic sample using an automated extraction and purification system includes optical tweezers; an input channel through which the sample is introduced; a chamber which receives the sample from the input channel; a collection channel through which selected particles of the sample are removed; and an output through which unselected particles of the sample are removed. At least one buffer input channel is provided. The input channel may allow sedimentation of the sample into the chamber by gravity. In another arrangement, the system includes an optical trapping system; a first channel through which the sample is introduced; a second channel through which buffer is introduced; wherein the optical tweezers are used to move selected particles of the sample from the first channel to the second channel. The selected particles may be sperm. The optical tweezers preferably utilize holographic optical trapping.

Abstract: The invention provides a method, apparatus and system for separating blood and other types of cellular components, and can be combined with holographic optical trapping manipulation or other forms of optical tweezing. One of the exemplary methods includes providing a first flow having a plurality of blood components; providing a second flow; contacting the first flow with the second flow to provide a first separation region; and differentially sedimenting a first blood cellular component of the plurality of blood components into the second flow while concurrently maintaining a second blood cellular component of the plurality of blood components in the first flow. The second flow having the first blood cellular component is then differentially removed from the first flow having the second blood cellular component. Holographic optical traps may also be utilized in conjunction with the various flows to move selected components from one flow to another, as part of or in addition to a separation stage.

Abstract: The invention provides a method and apparatus for separating blood into components, may be expanded to include other types of cellular components, and can be combined with holographic optical manipulation or other forms of optical tweezing. One of the exemplary methods includes providing a first flow having a plurality of blood components; providing a second flow; contacting the first flow with the second flow to provide a first separation region; and differentially sedimenting a first blood cellular component of the plurality of blood components into the second flow while concurrently maintaining a second blood cellular component of the plurality of blood components in the first flow. The second flow having the first blood cellular component is then differentially removed from the first flow having the second blood cellular component.