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.

Figure 6 from Turbo FISH: A Method for Rapid Single Molecule RNA FISH

Viral detection and identification in 20 min by rapid single-particle fluorescence in-situ hybridization of viral RNA

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

Frontiers Multiplexed Single-Cell in situ RNA Profiling

Publications — Raj Lab

PDF] Real-Time Monitoring of Fluorescence in Situ Hybridization Kinetics.

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

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RNA fluorescence in situ hybridization for high-content screening - ScienceDirect

Papers — Syd Shaffer Lab

Discriminating RNA variants with single-molecule allele-specific FISH - ScienceDirect

Quantitative gene expression analysis in Caenorhabditis elegans using single molecule RNA FISH.

FISH-Flow, a protocol for the concurrent detection of mRNA and protein in single cells using fluorescence in situ hybridization and flow cytometry

Intracellular Detection of Viral Transcription and Replication Using RNA FISH