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.

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

Real-Time Monitoring of Fluorescence in Situ Hybridization Kinetics

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

The Dynamics of mRNA Turnover Revealed by Single-Molecule Imaging in Single Cells - ScienceDirect

Stellaris RNA FISH Buffers LGC Biosearch Technologies

ClampFISH 2.0 enables rapid, scalable amplified RNA detection in situ

RNA Imaging with Multiplexed Error-Robust Fluorescence In Situ Hybridization (MERFISH).

Amplified Fluorescence in Situ Hybridization by Small and Bright Dye-Loaded Polymeric Nanoparticles

HT-smFISH – a cost-effective and flexible workflow for high-throughput single-molecule RNA imaging

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

RNA Imaging with Multiplexed Error-Robust Fluorescence In Situ Hybridization (MERFISH).

Biosensors, Free Full-Text

High-fidelity amplified FISH for the detection and allelic discrimination of single mRNA molecules