Turbo FISH: a method for rapid single molecule RNA FISH.
Advances in RNA fluorescence in situ hybridization (RNA FISH) have allowed practitioners to detect individual RNA molecules in single cells via fluorescence microscopy, enabling highly accurate and sensitive quantification of gene expression. However, current methods typically employ hybridization times on the order of 2-16 hours, limiting its potential in applications like rapid diagnostics. We present here a set of conditions for RNA FISH (dubbed Turbo RNA FISH) that allow us to make accurate measurements with no more than 5 minutes of hybridization time and 3 minutes of washing, and show that hybridization times can go as low as 30 seconds while still producing quantifiable images. We further show that rapid hybridization is compatible with our recently developed iceFISH and SNP FISH variants of RNA FISH that enable chromosome and single base discrimination, respectively. Our method is simple and cost effective, and has the potential to dramatically increase the throughput and realm of applicability of RNA FISH.
Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
Sydney M
SM
Shaffer
Min-Tzu
MT
Wu
Marshall J
MJ
Levesque
Arjun
A
Raj
D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D012313:RNA / Q000032:analysis* / Q000235:genetics
D012313:RNA