Abstract
Formalin-fixed paraffin-embedded (FFPE) tissues are typically the specimens available for FISH analysis of solid tissues, particularly of tumor specimens. Occasionally, tissue cores constructed as tissue microarrays from several patients are presented for simultaneous evaluation. FFPE sections can also be prepared from cell blocks derived from cell suspensions. The interphase fluorescence in situ hybridization assay employs specific nucleic acid sequences (probes) that target complementary sequences of interest to detect gains or losses of genes/gene loci or a fusion gene within the tissue. In this chapter, we describe the protocols utilized in our laboratory and include slide deparaffinization, pretreatment, protease treatment, hybridization, washing, and counterstaining. This protocol can be applied to all of the earlier FFPE preparations. In general, the assay takes 3 consecutive days to complete, although a more rapid assay can be performed.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Wolff AC, Hammond ME, Hicks DG et al (2007) American Society of Clinical Oncology/College of American Pathologists guideline recommendations for human epidermal growth factor receptor 2 testing in breast cancer. Arch Pathol Lab Med 131:18–43
Wolff AC, Hammond ME, Hicks DG et al (2013) Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update. J Clin Oncol 31:3997–4013. doi:10.1200/JCO.2013.50.9984
Petersen BL, Sorensen MC, Pedersen S et al (2004) Fluorescence in situ hybridization on formalin-fixed and paraffin-embedded tissue: optimizing the method. Appl Immunohistochem Mol Morphol 12(3):259–265
Müller S, Matthiesen SH, Nielsen KV (2009) Preparation of FFPE tissue slides for solid tumor FISH analysis. In: Kumar GL, Rudbeck L (eds) Education guide. Immunohistochemical (IHC) staining methods, 5th edn. Dako North America, Carpinteria, CA
Sommerlad C, Mehraein Y, Giersberg M et al (2002) Formalin-fixed and paraffin-embedded tissue sections. In: Rautenstrauss BW, Liehr T (eds) FISH technology. Springer, Berlin, Heidelberg
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Lim, A.S.T., Lim, T.H. (2017). Fluorescence In Situ Hybridization on Tissue Sections. In: Wan, T. (eds) Cancer Cytogenetics. Methods in Molecular Biology, vol 1541. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6703-2_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6703-2_11
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6701-8
Online ISBN: 978-1-4939-6703-2
eBook Packages: Springer Protocols