Hybridization Chain Reaction for mRNA Localization in Single Cells from Mouse and Human Cryosections.

ABSTRACT: In situ hybridization has been a robust method for detection of mRNA expression in whole-mount samples or tissue sections for more than 50 years. Recent technical advances for in situ hybridization have incorporated oligo-based probes that attain greater tissue penetration and signal amplification steps with restricted localization for visualization of specific mRNAs within single cells. One such method is third-generation in situ hybridization chain reaction (V3HCR). Here, we report an optimized protocol for V3HCR detection of gene expression using sectioned frozen tissues from mouse and human on microscope slides. Our methods and modifications for cryosectioning, tissue fixation, and processing over a three-day V3HCR protocol are detailed along with recommendations for aliquoting and sto

SUBMITTER: May-Zhang AA 

PROVIDER: S-EPMC9202517 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature