Analysis of the subcellular distribution of RNA by fluorescence in situ hybridization

In situ hybridization is a powerful method of examining the spatial distribution of RNA molecules at the subcellular level and serves as a basic technique in the fields of cell and developmental biology. In this technique, target RNAs are fixed in cells using formaldehyde and then hybridized to complementary probes labeled with modified nucleotides that are subsequently detected by immunohistochemical methods. Here, the procedures that are commonly used for the detection of RNA in mammalian tissues and cells are described, focusing on technical tips that improve the sensitivity, productivity, and reproducibility.