Acoustic Cytometry Methods and Protocols
First Claim
1. A method for analyzing bioparticles comprising:
- acoustically focusing one or more bioparticles through an interrogation zone;
optically exciting the one or more bioparticles in the interrogation zone with an excitation source;
detecting an optical signal from the bioparticles; and
analyzing the optical signal to characterize at least one quality or quantity parameter of the bioparticles.
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Abstract
Various embodiments disclosed herein comprise acoustic cytometry based methods, kits, computer software methods and systems to analyze a variety of bioparticles. In one embodiment, a method for analyzing bioparticles comprises: acoustically focusing one or more bioparticles through an interrogation zone; optically exciting the one or more bioparticles in the interrogation zone with an excitation source; detecting an optical signal from the bioparticles; and analyzing the optical signal to characterize at least one quality or quantity parameter of the bioparticles. Properties of biomolecules that may be analyzed include but are not limited to cell proliferation analysis, live/dead cell discrimination, cell cycle analysis, basic phenotyping, immunophenotyping, rare-event detection, apoptosis, phagocytosis, pinocytosis, detection of phosphoproteins, detection of one or more cellular markers, detection of one or more intracellular marker, detection of cancer cells, detection of pathological markers on a cell, microbial cell analysis and/or picophytoplankton analysis.
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Citations
37 Claims
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1. A method for analyzing bioparticles comprising:
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acoustically focusing one or more bioparticles through an interrogation zone; optically exciting the one or more bioparticles in the interrogation zone with an excitation source; detecting an optical signal from the bioparticles; and analyzing the optical signal to characterize at least one quality or quantity parameter of the bioparticles. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 28, 29, 30, 31, 32, 33)
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15. A method for detecting phosphoproteins on a cell disposed within a fluid medium, comprising:
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stimulating or inhibiting the cell with a kinase or a kinase inhibitor respectively to phorsporylate or de-phosphorylate one or more proteins on the cell; contacting the cell with one or more antibody specific to detect the one or more phosphorylated protein; acoustically focusing the cell in the fluid medium; optically exciting the cell with an excitation source; detecting an optical signal from the cell; and analyzing the optical signal, wherein the optical signal is indicative of the presence or absence of the one or more phosphorylated protein.
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16. A method for detecting fluorescent protein expression on a cell disposed within a fluid medium, comprising:
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transfecting the cell with one or more fluorescent proteins; acoustically focusing the cell in the fluid medium; optically exciting the cell with an excitation source; detecting one or more optical signals from the cell; and analyzing the optical signal, wherein the detection of an optical signal corresponding to one or more fluorescent protein is indicative of the presence of expression of the one or more fluorescent proteins and the absence of an optical signal corresponding to one or more fluorescent protein is indicative of the absence of expression of the fluorescent protein. - View Dependent Claims (17, 18, 19, 20)
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21. A method for detection a rare event within a population of cells, the method comprising:
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acoustically focusing the population of cells; optically exciting the population of cells with an excitation source; detecting one or more optical signals from the population of cells; and analyzing the optical signal, wherein the detection of an optical signal corresponding to a rare event is indicative of the presence of the rare event and the absence of an optical signal corresponding to a rare event is indicative of the absence of the rare event. - View Dependent Claims (22, 23, 24, 25, 26, 27)
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34. A method for detecting cell apoptosis, the method comprising:
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acoustically focusing one or more cells disposed within a fluid; optically exciting the one or more cells with an excitation source; detecting one or more optical signals from the cells; and analyzing the detected optical signals to identify morphological or biochemical changes that are indicative of cell apoptosis. - View Dependent Claims (35, 36, 37)
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Specification