Four-Color STED Super-Resolution RNA Fluorescent In Situ Hybridization and Immunocytochemistry to Visualize Mitochondrial mRNAs in Context with Mitochondrial Ribosomes

Methods Mol Biol. 2023:2661:303-316. doi: 10.1007/978-1-0716-3171-3_17.

Abstract

High-resolution imaging has enabled scientists to explore the mitochondrial network at remarkable resolution. This has been exploited to help increase our knowledge of how mitochondrial gene expression is compartmentalized in cultured cells. Here, we provide detailed methodology to simultaneously visualize up to four components including mtDNA-encoded transcripts, submitochondrial marker proteins, mitoribosomal subunits, or core members of the translational apparatus using STED super-resolution nanoscopy.

Keywords: Mitochondrial mRNA; RNA in situ hybridization; STED super-resolution; Super-resolution nanoscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescent Dyes
  • Immunohistochemistry
  • In Situ Hybridization, Fluorescence
  • Microscopy, Fluorescence / methods
  • Mitochondrial Ribosomes*
  • RNA* / genetics

Substances

  • RNA
  • Fluorescent Dyes