Double fluorescence in situ hybridization in fresh brain sections.
Here we describe a modified version of a double fluorescence in situ hybridization (dFISH) method optimized for detecting two mRNAs of interest in fresh frozen brain sections. Our group has successfully used this approach to study gene co-regulation. More specifically, we have used this dFISH method to explore the anatomical organization, neurochemical properties, and the impact of sensory experience in central sensory circuits, at single cell resolution. This protocol has been validated in brain tissue from mice, rats and songbirds but is expected to be easily adaptable to other vertebrate species, as well as to an array of non-neural tissues. In this film we provide a detailed demonstration of the main steps of this procedure.
Jeong
Jin Kwon
JK
Department of Brain and Cognitive Sciences, University of Rochester, USA.
Chen
Zhuoxun
Z
Tremere
Liisa A
LA
Pinaud
Raphael
R
D000818:Animals; D001923:Brain Chemistry; D005629:Frozen Sections / Q000379:methods; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D051379:Mice; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D020308:Songbirds
D012333:RNA, Messenger