Optimization and evaluation of fluorescence in situ hybridization chain reaction in cleared fresh-frozen brain tissues.

ABSTRACT: Transcript labeling in intact tissues using in situ hybridization chain reaction has potential to provide vital spatiotemporal information for molecular characterization of heterogeneous neuronal populations. However, large tissue labeling in non-perfused or fresh-frozen rodent and postmortem human samples, which provide more flexible utilization than perfused tissues, is largely unexplored. In the present study, we optimized the combination of in situ hybridization chain reaction in fresh-frozen rodent brains and then evaluated the uniformity of neuronal labeling between two clearing methods, CLARITY and iDISCO⁺. We found that CLARITY yielded higher signal-to-noise ratios but more limited imaging depth and required longer clearing times, whereas, iDISCO⁺ resulted in

SUBMITTER: Kumar V 

PROVIDER: S-EPMC7962668 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature