Simultaneous detection of rnas and proteins with subcellular resolution

Sunjong Kwon, Koei Chin, Michel Nederlof

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

We describe the detailed methods of “immunoFISH” to analyze the expression level and the spatial localization of RNA transcripts and proteins on cultured cells and formal-fixed, paraffin-embedded (FFPE) tissue sections. On cultured cells, we detect specific transcripts using the Stellaris fluorescence in situ hybridization (FISH) probes labeled with fluorophores that target multiple regions along the desired transcripts and proteins combining the immunofluorescent staining. On FFPE tissue sections, we use the RNAscope FISH probes, modified branched DNA (bDNA) probes to amplify the RNA signals, followed by immunofluorescent staining for protein detection. The abundance, composition, and spatial distribution are determined by signals from fluorescently labeled proteins and individual transcripts of images acquired using high-resolution fluorescence microscopy.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages59-73
Number of pages15
DOIs
StatePublished - 2020

Publication series

NameMethods in Molecular Biology
Volume2161
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Immunofluorescent
  • Subcellular localization
  • immunoFISH
  • smFISH

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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