Methods and apparatus for discovering, identifying and comparing biological activity mechanisms
First Claim
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1. A method of identifying the biological mechanisms affected by a selected gene, comprising a) culturing a first reference cell under reproducible conditions;
- b) processing the first reference cell through an assay in the presence of a perturbation;
c) collecting one or more images of the first cell to detect a first cell assay response to the respective perturbation;
d) culturing a second cell under the reproducible conditions of step a), wherein the first reference cell and the second test-cell are the same cell species, and the second test-cell is altered to modify the expression of the protein encoded by the selected gene;
e) processing the second test-cell through the assay of step b) in the presence of the same perturbation;
f) collecting one or more images of the second cell to detect a second test-cell assay response to the respective perturbation;
g) comparing the one or more images obtained of the first reference cell to the one or more images obtained of the second altered test-cell to identify assay response image changes between the first reference cell and the second test-cell, wherein the assay response image changes correspond to the biological mechanisms affected by the selected gene.
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Abstract
Provided herein are methods and devices for the assessment and identification of cellular biological activity mechanisms; the assessment and identification of the changes in cellular biological activity mechanisms caused by cellular perturbations; the assessment and identification of the cellular function, or biological activity mechanisms of genes and gene products and; and the identification of the many genes and their products that collectively act together in a biological mechanism.
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Citations
52 Claims
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1. A method of identifying the biological mechanisms affected by a selected gene, comprising
a) culturing a first reference cell under reproducible conditions; -
b) processing the first reference cell through an assay in the presence of a perturbation;
c) collecting one or more images of the first cell to detect a first cell assay response to the respective perturbation;
d) culturing a second cell under the reproducible conditions of step a), wherein the first reference cell and the second test-cell are the same cell species, and the second test-cell is altered to modify the expression of the protein encoded by the selected gene;
e) processing the second test-cell through the assay of step b) in the presence of the same perturbation;
f) collecting one or more images of the second cell to detect a second test-cell assay response to the respective perturbation;
g) comparing the one or more images obtained of the first reference cell to the one or more images obtained of the second altered test-cell to identify assay response image changes between the first reference cell and the second test-cell, wherein the assay response image changes correspond to the biological mechanisms affected by the selected gene. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 51)
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28. A method of producing a fingerprint of assay responses caused by a perturbation, comprising
a) culturing a first reference cell under reproducible conditions; -
b) processing the first reference cell through a multiplicity of assay experiments in the absence of a perturbation;
c) collecting one or more images of the first reference cell to detect a first cell assay response to the respective assays;
d) culturing a second test-cell under the reproducible conditions of step a), wherein the first reference cell and the second test-cell are the same cell species;
e) processing the second test-cell through the same multiplicity of assay experiments of step b) in the presence of a perturbation;
f) collecting one or more images of the second test-cell to detect a second test-cell assay response to the respective perturbation;
g) comparing the one or more images obtained of the first reference cell to the one or more images obtained of the second test-cell to identify assay response image changes between the first reference cell and the second test-cell, wherein the assay response image changes correspond to a fingerprint of assay responses caused by the perturbation. - View Dependent Claims (29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52)
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Specification