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.

Genome oligopaint via local denaturation fluorescence in situ hybridization - ScienceDirect

Methods for spatial and temporal imaging of the different steps involved in RNA processing at single‐molecule resolution - Markey - 2021 - WIREs RNA - Wiley Online Library

Using Single Molecule mRNA Fluorescent in Situ Hybridization (RNA-FISH) to Quantify mRNAs in Individual Murine Oocytes and Embryos

Methods for spatial and temporal imaging of the different steps involved in RNA processing at single‐molecule resolution - Markey - 2021 - WIREs RNA - Wiley Online Library

PDF] A technical review and guide to RNA fluorescence in situ hybridization

Genome oligopaint via local denaturation fluorescence in situ hybridization - ScienceDirect

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