A robust method for isotopic riboprobe in situ hybridisation to localise mRNAs in routine pathology specimens

Eur J Histochem. 1998;42(2):121-32.


In situ Hybridisation (ISH) to detect mRNA is widely applicable to studies of human pathology and experimental models including transgenic mice, where it can provide crucial information as to where a gene is expressed, that is not available in other ways. ISH was used to establish that relatively high levels of expression of E-cadherin mRNA were associated with long-term survival in colorectal cancer, before suitable antisera became available. There is increasing awareness that ISH can be used to help select between candidate disease genes found at a linked locus, and that ISH can help validate therapeutic targets. For instance, when several closely related receptor genes are expressed in a tissue, it may be possible to determine which combinations are expressed together in a single cell type. These diverse applications demand a robust method that works on a variety of clinical and experimental materials, and is easily interpreted. To date, the ICRF in situ Hybridisation Service has hybridised over 30,000 sections of principally formalin-fixed, paraffin-embedded tissues, with a high success rate. The practicalities of our preferred method are discussed and key steps for quality control highlighted.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Automation
  • Humans
  • In Situ Hybridization / methods*
  • Isotope Labeling
  • RNA Probes
  • RNA, Messenger / analysis*


  • RNA Probes
  • RNA, Messenger