Double fluorescence in situ hybridization in fresh brain sections
Journal of Visualized Experiments
Biotin; Digoxigenin; Double FISH (dFISH); Fluorescence in situ hybridization (FISH); Issue 42; Neuroscience; Non-radioactive riboprobes; Vertebrate brain
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. © JoVE 2006-2011 All Rights Reserved.
Jeong, J., Chen, Z., Tremere, L., & Pinaud, R. (2010). Double fluorescence in situ hybridization in fresh brain sections. Journal of Visualized Experiments, (42). http://dx.doi.org/10.3791/2102