Double fluorescence in situ hybridization in fresh brain sections

Document Type

Journal Article

Publication Date

1-1-2010

Journal

Journal of Visualized Experiments

Issue

42

DOI

10.3791/2102

Keywords

Biotin; Digoxigenin; Double FISH (dFISH); Fluorescence in situ hybridization (FISH); Issue 42; Neuroscience; Non-radioactive riboprobes; Vertebrate brain

Abstract

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.

This document is currently not available here.

Share

COinS