title: A robust method for isotopic riboprobe in situ hybridisation to localise mRNAs in routine pathology specimens.
abstract: 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.
authors.affiliation: In Situ Hybridisation Service, Imperial Cancer Research Fund, London, UK. poulsom@icrf.icnet.uk
authors.forename: R
authors.initials: R
authors.lastname: Poulsom
authors.forename: J M
authors.initials: JM
authors.lastname: Longcroft
authors.forename: R E
authors.initials: RE
authors.lastname: Jeffery
authors.forename: L A
authors.initials: LA
authors.lastname: Rogers
authors.forename: J H
authors.initials: JH
authors.lastname: Steel
mesh_terms: D001331:Automation; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D007553:Isotope Labeling; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis*
chemical_list: D015347:RNA Probes; D012333:RNA, Messenger