Abstract:
The present invention relates to a method of processing forensic trace or touch DNA evidence in order to differentially label cells, and then isolate them into distinct fractions which can be processed using standard downstream DNA profiling methods. A mixture of cells and/or cellular components in fluid suspension are first labeled with one or more markers which bind specifically to genetic sequences of interest, and which can be detected optically. The labeled cells/nuclei are placed into a fluidic cartridge, and are transported via fluid flow to an inspection and separation region, observed and identified according to their labels using optical microscopy, and are moved to output channels corresponding to the different labels, degree of labeling, or combination of labels, and to a different subset of possible genetic profiles. The separated cell fractions are extracted from the cartridge via their respective outputs, and can be genetically profiled via short- tandem-repeat (STR) analysis.
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:
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. 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), allowing for more efficient detection and reduced test time. The 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. An 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. Additional methods and devices for target capturing include a substantially planar surface having an optimized angle for capture, or alternative solid phase geometries for capture.
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:
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. 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:
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.