Sequential analysis of biological samples
First Claim
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1. A method for in situ detection of a plurality of targets in a biological sample, comprising:
- (a) contacting the biological sample in situ with a plurality of target-binding probes to form a plurality of target-bound probes;
(b) detecting a first set of fluorescence signals from a first set of the plurality of target-bound probes;
(c) modifying the detected fluorescence signals using a chemical agent;
(d) generating a second set of fluorescence signals after step (c) from a second set of the plurality of target-bound probes;
(e) detecting the second set of fluorescence signals; and
(f) repeating steps (c) to (e) multiple times with a third, a fourth, or an nth set of the plurality of target-bound probes to detect a third, a fourth, or an nth set of fluorescence signals.(f) repeating steps (c) to (e) multiple times with a third, a fourth, or an nth set of the plurality of target-bound probes to detect a third, a fourth, or an nth set of fluorescence signals.
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Abstract
Methods for detecting a plurality of targets in a biological sample are provided. The method comprises contacting the biological sample with a plurality of target-binding probes simultaneously to form a plurality of target-bound probes and observing the signals from the target-bound probes sequentially. An associated kit and device for detection of the plurality of targets are also provided.
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Citations
23 Claims
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1. A method for in situ detection of a plurality of targets in a biological sample, comprising:
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(a) contacting the biological sample in situ with a plurality of target-binding probes to form a plurality of target-bound probes; (b) detecting a first set of fluorescence signals from a first set of the plurality of target-bound probes; (c) modifying the detected fluorescence signals using a chemical agent; (d) generating a second set of fluorescence signals after step (c) from a second set of the plurality of target-bound probes; (e) detecting the second set of fluorescence signals; and (f) repeating steps (c) to (e) multiple times with a third, a fourth, or an nth set of the plurality of target-bound probes to detect a third, a fourth, or an nth set of fluorescence signals. (f) repeating steps (c) to (e) multiple times with a third, a fourth, or an nth set of the plurality of target-bound probes to detect a third, a fourth, or an nth set of fluorescence signals. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method for in situ detection of a plurality of targets in a biological sample, comprising:
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(a) contacting the biological sample in situ with a plurality of target-binding probes to form a plurality of target-bound probes; (b) generating a first set of fluorescence signals from a first set of the plurality of target-bound probes; (c) detecting the first set of fluorescence signals; (d) modifying the detected fluorescence signals after step (c) using a chemical agent; (e) generating a second set of fluorescence signals after step (d) from a second set of the plurality of target-bound probes; (f) detecting the second set of fluorescence signals; and (g) repeating steps (d) to (f) multiple times with a third, a fourth, or an nth set of the plurality of target-bound probes to detect a third, a fourth, or an nth set of fluorescence signals. - View Dependent Claims (16, 17, 18, 19)
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20. A method for in situ detection of a plurality of targets in a tissue section, comprising:
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(a) contacting the tissue section in situ with a plurality of target-binding probes to form a plurality of target-bound probes; (b) contacting the plurality of target-bound probes with a first set of fluorescent probes that are capable of binding a first set of the plurality of target-bound probes, and are capable of generating a first set of fluorescence signals; (c) detecting the first set of the plurality of targets via observing the first set of fluorescence signals; (d) modifying the detected first set of fluorescence signals by applying a chemical agent; and (e) repeating steps (b) to (d) multiple times after step (d) using a subsequent set of fluorescent probes to detect a subsequent set of the plurality of targets, wherein the subsequent set of fluorescent probes are capable of binding a subsequent set of the plurality of target-bound probes, and are capable of generating a subsequent set of fluorescence signals. - View Dependent Claims (21, 22, 23)
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Specification