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Methods Mol Biol. 2018;1767:385-393. doi: 10.1007/978-1-4939-7774-1_21.

Cell-to-Cell Transcription Variability as Measured by Single-Molecule RNA FISH to Detect Epigenetic State Switching.

Author information

1
Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
2
Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands. p.j.verschure@uva.nl.

Abstract

Single-molecule RNA fluorescent in situ hybridization (smRNA FISH) allows for the visualization, localization, and quantification of RNA transcripts within individual cells and tissues using custom-designed fluorescently labeled oligonucleotide probes. Here we describe a protocol for the preparation, imaging, and analysis of a smRNA FISH experiment that can be applied to any RNA of choice. We also provide insights as to how this powerful tool can be used to study epigenetic regulation, for example, following the epigenetic editing of genes.

KEYWORDS:

Epigenetics; Fixation; Fluorescence microscopy; Hybridization; Image analysis; Permeabilization; RNA FISH

PMID:
29524147
DOI:
10.1007/978-1-4939-7774-1_21
[Indexed for MEDLINE]

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