Triple-Labeling Whole-Mount In Situ Hybridization Method for Analysis of Overlapping Gene Expression in Brain Tissue with High Level of Autofluorescence

Abstract

Background: Detection of multiple co-localized mRNAs by conventional chromogenic in situ hybridization is difficult because the first reaction product can obscure subsequent ones. Multi-color fluorescent in situ hybridization (FISH) offers simultaneous detection of multiple mRNAs but has low sensitivity and in cells of the central nervous system (CNS), is hampered by high background autofluorescence.