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.

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

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

IJMS, Free Full-Text

Intracellular Detection of Viral Transcription and Replication Using RNA FISH

Detection of mRNA Transfer Between Mammalian Cells in Coculture by Single- Molecule Fluorescent In Situ Hybridization (smFISH)

Papers — Syd Shaffer Lab

FISHing for Accurate Transcript Counts with Single-Molecule FISH

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

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

Use of Split FISH for Spatial Transcriptomics- Oxford Instruments

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

Multiplexed detection of SARS-CoV-2 genomic and subgenomic RNA using in situ hybridization

FISHing fish - the Node

Strength in numbers: quantitative single‐molecule RNA detection assays - Gaspar - 2015 - WIREs Developmental Biology - Wiley Online Library