Photoactivated chemical bleaching of dyes
First Claim
1. A method of probing multiple targets in a biological sample comprising:
- (a) providing a biological sample containing multiple targets;
(b) binding multiple probes to multiple targets present in the biological sample and wherein said probe comprises a binder and at least one of an optical signal generator or a fluorescent signal generator wherein the binder is capable of binding specifically to one or more of the targets present in the biological sample;
(c) observing a first set of signals from the first set of probes bound in step (b);
(d) contacting the sample comprising the bound probe of step (b) with an electron transfer reagent wherein the electron transfer reagent is a borate salt represented by the following structural formula;
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Abstract
Methods comprising the use of photoactivated chemical bleaching for detecting multiple targets in a biological sample are provided. The methods include the steps of providing a biological sample containing multiple targets, binding at least one probe to one or more target present in the sample, and observing a signal from the probe. The method further includes the steps of contacting the sample comprising the bound probe with an electron transfer reagent and irradiating the sample, thereby initiating a photoreaction that substantially inactivates the probe by photoactivated chemical bleaching. The method further includes the steps of binding at least one probe to one or more target present in the sample, and observing a signal from the probe. The process of binding, observing and bleaching may be iteratively repeated.
29 Citations
7 Claims
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1. A method of probing multiple targets in a biological sample comprising:
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(a) providing a biological sample containing multiple targets; (b) binding multiple probes to multiple targets present in the biological sample and wherein said probe comprises a binder and at least one of an optical signal generator or a fluorescent signal generator wherein the binder is capable of binding specifically to one or more of the targets present in the biological sample; (c) observing a first set of signals from the first set of probes bound in step (b); (d) contacting the sample comprising the bound probe of step (b) with an electron transfer reagent wherein the electron transfer reagent is a borate salt represented by the following structural formula; - View Dependent Claims (2, 3, 4)
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5. A high throughput multiplexing biological sample analysis method, the method comprising:
a multiple signal cycling process comprising staining and imaging of the biological sample in each cycle of the multiple signal cycling process, wherein in each of the cycle, staining and imaging of the biological sample is followed by applying an electron transfer reagent and irradiation of the biological sample wherein the electron transfer reagent is a borate salt represented by the following structural formula; - View Dependent Claims (6)
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7. A series of at least two images depicting optically labeled biological targets wherein:
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the images are obtained in the process of probing multiple targets in a biological sample, wherein the process comprises; (a) providing a biological sample containing multiple targets; (b) binding at least one optical probe to one or more targets present in the biological sample, wherein the optical probe comprises a binder conjugated to an optical signal generator and wherein the binder is capable of binding specifically to the one or more targets present in the biological sample; (c) observing a signal from the optical probe bound in step (b); (d) contacting the sample comprising the bound optical probe of step (b) with an electron transfer reagent wherein the electron transfer reagent is a borate salt represented by the following structural formula;
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Specification