Expression analysis of mRNA in formalin-fixed, paraffin-embedded archival tissues by mRNA in situ hybridization

Methods : a Companion to Methods in Enzymology
Ralf T HenkeAnton Wellstein

Abstract

Gene expression in diseased tissues can indicate the contribution to a disease process and potentially guide therapeutic decision-making. Archival tissues with associated clinical outcome may be useful to discover or validate the role of a candidate gene in a disease process or the response to therapy. Such archival tissues are commonly formalin-fixed and paraffin-embedded, restricting the methods available for gene expression analysis. Obviously, the detection of proteins in tissues requires adaptation for each protein and the detection of secreted proteins can prove difficult or of reduced value since the protein detected may not reflect the total amount produced. Thus, we describe here a reliable method for the detection of mRNA in archival tissues. The method for mRNA in situ hybridization (ISH) was adapted by us for >15 different genes and applied to several hundred tissue microarrays (TMAs) and full sections generating >10,000 expression data points. We also discuss the utility of TMAs to simultaneously analyze several hundred tissue samples on one slide to minimize variability and preserve valuable tissue samples. Experimental protocols are provided that can be implemented without major hurdles in a typical molecular pat...Continue Reading

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Citations

May 28, 2009·Proceedings of the National Academy of Sciences of the United States of America·Verónica A BurzioLuis O Burzio
May 7, 2009·Breast Cancer Research and Treatment·Tyler LahusenAnna T Riegel
May 28, 2013·Genome Medicine·Jeremy A MillerDaniel H Geschwind
Jul 30, 2011·Diseases of Aquatic Organisms·L Al-Hussinee, J S Lumsden
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Mar 8, 2013·Clinical Biochemistry·Nils Becker, Christina M Lockwood
Jun 8, 2007·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Elena TassiAnton Wellstein
Apr 28, 2020·The Biochemical Journal·Anna Wilbrey-ClarkSarah A Teichmann
Dec 20, 2019·Nucleic Acids Research·Lena Voith von VoithenbergGovind V Kaigala
May 13, 2009·Proceedings of the National Academy of Sciences of the United States of America·Krissa A GibbyAnton Wellstein

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