Photoactivated chemical bleaching of dyes using borates

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 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 a borate compound of Formula I;
• R1R2R3R4BXa I
• where each R1, R2, and R3 and R4 is, independently, an alkyl, an alkylaryl, an arylalkyl, or an aryl, with the proviso that at least one of R1, R2, and R3 and R4 is an alkyl group and at least one of R1, R2, and R3 and R4 group is positively charged;
• X is an anion;
• and
• a equals 0, 1, 2, or 3;
• (e) irradiating the sample of step (d);
• (f) generating a second set of signals from the second set of probes bound in step (b);
• (g) observing the second set of signals.

A series of at least two images depicting optically labeled biological targets wherein:

• 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 sample;
• (c) capturing a first image a signal from the optical probe bound in step (b)
• (d) contacting the sample comprising the bound optical probe of step (b) with a borate compound of Formula I;
• R1R2R3R4BXa I
• where each R1, R2, and R3 and R4 is, independently, an alkyl, an alkylaryl, an arylalkyl, or an aryl, with the proviso that at least one of R1, R2, and R3 and R4 is an alkyl group and at least one of R1, R2, and R3 and R4 group is positively charged;
• X is an anion;
• and
• a equals 0, 1, 2, or 3;
• (e) irradiating the sample of step (d);
• (f) binding at least one optical probe to one or more targets present in the sample of step (e);
• and
• (g) capturing a subsequent image a signal from the optical probe bound in step (f).