Multiplexed miRNA fluorescence in situ hybridization for formalin-fixed paraffin-embedded tissues

Methods Mol Biol. 2014:1211:171-87. doi: 10.1007/978-1-4939-1459-3_14.

Abstract

Multiplexed miRNA fluorescence in situ hybridization (miRNA FISH) is an advanced method for visualizing differentially expressed miRNAs, together with other reference RNAs, in archival tissues. Some miRNAs are excellent disease biomarkers due to their abundance and cell-type specificity. However, these short RNA molecules are difficult to visualize due to loss by diffusion, probe mishybridization, and signal detection and signal amplification issues. Here, we describe a reliable and adjustable method for visualizing and normalizing miRNA signals in formalin-fixed paraffin-embedded (FFPE) tissue sections.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescent Dyes / analysis
  • Formaldehyde / chemistry
  • Humans
  • In Situ Hybridization, Fluorescence / methods*
  • MicroRNAs / analysis*
  • Microscopy, Fluorescence / methods
  • Paraffin Embedding / methods
  • RNA Probes / analysis
  • Tissue Fixation / methods

Substances

  • Fluorescent Dyes
  • MicroRNAs
  • RNA Probes
  • Formaldehyde