Detecting microRNA in human cancer tissues with fluorescence in situ hybridization

Methods Mol Biol. 2013:1039:19-27. doi: 10.1007/978-1-62703-535-4_2.

Abstract

The technique of nucleic acid in situ hybridization is an effective method for identifying the existence and abundance of nucleic acids in tissue sections or cytological preparations. Such a method has the advantage of keeping morphological relationships intact while identifying changes at the molecular level. As a noncoding regulatory RNA, microRNA has been found to intricately control many physiological and pathological conditions. We provide here a representative fluorescence in situ hybridization protocol for microRNA detection, and note commonly used alternatives, and some troubleshooting points. The method described is based on formalin-fixed paraffin-embedded oral cancer tissues but should be broadly applicable to similarly processed tissues of other types of cancer.

MeSH terms

  • Humans
  • In Situ Hybridization, Fluorescence / methods*
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Neoplasms / genetics*
  • Neoplasms / metabolism
  • Paraffin Embedding
  • Tissue Fixation / methods

Substances

  • MicroRNAs