Sequential analysis of biological samples
First Claim
1. A kit for detection of multiple targets in a biological sample, comprising:
- multiple probes, wherein each probe comprises a binder and a fluorophore, wherein each probe is configured to specifically bind to a target via the binder, wherein the binder comprises a primary antibody and wherein the fluorophore is coupled to a secondary antibody that binds to the target via the primary antibody; and
at least one chemical agent in a concentration that, when applied to each probe, is configured to modify the fluorescence signal provided by the fluorophore of at least one probe without affecting the integrity of the binder in the at least one probe such that the primary antibody, when bound to the target, remains bound to the target after modification of the fluorescent signal by the at least one chemical agent under conditions in which the fluorophore is destroyed.
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Accused Products
Abstract
Methods for detecting multiple targets in a biological sample are provided. The methods includes contacting the sample with a first probe; physically binding the first probe to a first target; observing a first signal from the first probe; applying a chemical agent to modify the first signal; contacting the sample with a second probe; physically binding the second probe to a second target; and observing a second signal from the second probe. The methods disclosed herein also provide for multiple iterations of binding, observing, signal modification for deriving information about multiple targets in a single sample. An associated kit and device are also provided.
32 Citations
25 Claims
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1. A kit for detection of multiple targets in a biological sample, comprising:
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multiple probes, wherein each probe comprises a binder and a fluorophore, wherein each probe is configured to specifically bind to a target via the binder, wherein the binder comprises a primary antibody and wherein the fluorophore is coupled to a secondary antibody that binds to the target via the primary antibody; and at least one chemical agent in a concentration that, when applied to each probe, is configured to modify the fluorescence signal provided by the fluorophore of at least one probe without affecting the integrity of the binder in the at least one probe such that the primary antibody, when bound to the target, remains bound to the target after modification of the fluorescent signal by the at least one chemical agent under conditions in which the fluorophore is destroyed. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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12. A kit for detection of multiple targets in a biological sample, comprising:
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a first probe, comprising a first binder and a first fluorophore, wherein the first probe is configured to specifically bind to a first target via the first binder, and wherein the first fluorophore is configured to provide a first fluorescence signal; a second probe, comprising a second binder and a second fluorophore, wherein the second probe is configured to specifically bind to a second target via the second binder, and wherein the second fluorophore is configured to provide a second fluorescence signal; a first chemical agent in a concentration that, when applied to each probe, is configured to modify the first fluorescence signal provided by the first fluorophore without affecting the integrity of the first binder such that the first binder, when bound to the first target, remains bound to the first target after modification of the first fluorescent signal by the first chemical agent under conditions in which the first fluorophore is destroyed; and a second chemical agent configured to modify the second fluorescence signal provided by the second fluorophore without affecting the integrity of the second binder such that the second binder, when bound to the second target, remains bound to the second target after modification of the second fluorescent signal by the second chemical agent. - View Dependent Claims (13, 14, 15)
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16. A kit for detection of multiple proteins in a single tissue section, comprising:
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a plurality of cyanine dye-labeled antibodies; and at least one oxidizing agent in a concentration that, when applied to the cyanine dye-labeled antibodies, is configured to modify the cyanine fluorescence of at least one of the plurality of cyanine dye-labeled antibodies without affecting the integrity of the antibodies.
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Specification