Methods of analyzing an H and E stained biological sample

摘要

Methods comprising the use probing multiple targets in a H&E stained biological sample are provided. The methods include the steps of providing a hematoxylin and eosin stained biological sample containing multiple targets, observing the sample, removing the hematoxylin and partially removing the eosin by washing the sample, contacting the sample with a borate salt, and irradiating the sample to remove the residual eosin fluorescence. The method further includes the optionally performing the additional steps of binding at least one probe to one or more targets to the sample, observing a signal from the probe and contacting the sample with a bleaching agent. The process of binding, observing and bleaching may be iteratively repeated.

主权项

1. A sequential method of preparing a hematoxylin and eosin stained biological sample for analysis comprising:
(a) providing a hematoxylin and eosin stained biological sample containing multiple targets; (b) observing the hematoxylin and eosin si al from the stained biological sample; (c) removing the hematoxylin and partially removing the eosin by washing the sample; (d) contacting the sample with an electron transfer reagent where the electron transfer reagent is a borate salt represented by the following structural formula: wherein:each R1, R2, and R3 is, independently, an alkyl, an alkenyl, an akynyl, an aryl or a heteroaryl, wherein said alkyl, alkenyl, alkynyl, aryl or heteroaryl is optionally substituted with one or more substituents selected from the group consisting of (C1-C4)alkyl, (C1-C4)alkoxy, (C1-C4)alkylamino, amino, hydroxyl, cyano, halogen, or nitro,R4 is an alkyl, an alkenyl, or an akynyl, wherein said alkyl, alkenyl, or alkynyl is optionally substituted with one or more substituents selected from the group consisting of (C1-C4)alkyl, aryl, (C1-C4)alkoxy, (C1-C4)alkylamino, amino, hydroxyl, cyano, halogen, or nitro, andM+ is selected from the group consisting of organic and inorganic cations; (e) irradiating the sample of step (d) to remove residual fluorescence.