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Fluorescence In Situ Hybridization of Cells, Chromosomes, and Formalin-Fixed Paraffin-Embedded Tissues

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Molecular Profiling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1606))

Abstract

Fluorescence in situ hybridization (FISH) with DNA probes allows the visualization of gene copy number and localization of specific DNA targets with fluorescence microscopy. Cells in culture, metaphase chromosomes, and tissue sections are fixed and prepared on glass slides. Both the DNA in the cells and fluorescently labeled probe are denatured, and the labeled probe is allowed to hybridize to the cellular DNA. The slides are washed, counterstained, and viewed via fluorescence microscopy. We describe the basic method for preparing slides and probes for studies involving DNA copy number changes and structural chromosome rearrangements in formalin-fixed paraffin-embedded (FFPE) tissue sections and cell culture preparations.

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References

  1. Kallioniemi A, Visakorpi T, Karhu R, Pinkel D, Kallioniemi OP (1996) Gene copy number analysis by fluorescence in situ hybridization and comparative genomic hybridization. Methods 9(1):113–121

    Article  CAS  PubMed  Google Scholar 

  2. Pinkel D, Gray JW, Trask B, van den Engh G, Fuscoe J, van Dekken H (1986) Cytogenetic analysis by in situ hybridization with fluorescently labeled nucleic acid probes. Cold Spring Harb Symp Quant Biol 51(Pt 1):151–157

    Article  CAS  PubMed  Google Scholar 

  3. Waye JS, Willard HF (1985) Chromosome-specific alpha satellite DNA: nucleotide sequence analysis of the 2.0 kilobasepair repeat from the human X chromosome. Nucleic Acids Res 13(8):2731–2743

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Cajulis RS, Frias-Hidvegi D, Yu GH, Eggena S (1996) Detection of numerical chromosomal abnormalities by fluorescence in situ hybridization of interphase cell nuclei with chromosome-specific probes on archival cytologic samples. Diagn Cytopathol 14(2):178–181. doi:10.1002/(SICI)1097-0339(199603)14:2<178::AID-DC14>3.0.CO;2-J

  5. Cremer T, Lichter P, Borden J, Ward DC, Manuelidis L (1988) Detection of chromosome aberrations in metaphase and interphase tumor cells by in situ hybridization using chromosome-specific library probes. Hum Genet 80(3):235–246

    Article  CAS  PubMed  Google Scholar 

  6. Mayer JA, Pham T, Wong KL, Scoggin J, Sales EV, Clarin T, Pircher TJ, Mikolajczyk SD, Cotter PD, Bischoff FZ (2011) FISH-based determination of HER2 status in circulating tumor cells isolated with the microfluidic CEE platform. Cancer Genet 204(11):589–595. doi:10.1016/j.cancergen.2011.10.011

    Article  CAS  PubMed  Google Scholar 

  7. Pinkel D, Albertson DG (2005) Array comparative genomic hybridization and its applications in cancer. Nat Genet 37(Suppl):S11–S17. doi:10.1038/ng1569

    Article  CAS  PubMed  Google Scholar 

  8. Ventura RA, Martin-Subero JI, Jones M, McParland J, Gesk S, Mason DY, Siebert R (2006) FISH analysis for the detection of lymphoma-associated chromosomal abnormalities in routine paraffin-embedded tissue. J Mol Diagn 8(2):141–151. doi:10.2353/jmoldx.2006.050083

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Wang N (2002) Methodologies in cancer cytogenetics and molecular cytogenetics. Am J Med Genet 115(3):118–124. doi:10.1002/ajmg.10687

    Article  PubMed  Google Scholar 

  10. Blancato JK, Williams MS, Dickson RB (2006) Fluorescence in situ hybridization assessment of c-myc gene amplification in breast tumor tissues. Methods Mol Med 120:297–307

    CAS  PubMed  Google Scholar 

  11. Chin SF, Daigo Y, Huang HE, Iyer NG, Callagy G, Kranjac T, Gonzalez M, Sangan T, Earl H, Caldas C (2003) A simple and reliable pretreatment protocol facilitates fluorescent in situ hybridisation on tissue microarrays of paraffin wax embedded tumour samples. Mol Pathol 56(5):275–279

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Saraiva NZ, Perecin F, Meo SC, Ferreira CR, Tetzner TA, Garcia JM (2009) Demecolcine effects on microtubule kinetics and on chemically assisted enucleation of bovine oocytes. Cloning Stem Cells 11(1):141–152. doi:10.1089/clo.2008.0044. http://luckylion.de/vasco/tutorials/indesignthesis/mol_toolbox.php

    Article  CAS  PubMed  Google Scholar 

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Author information

Authors and Affiliations

  1. Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Washington, DC, USA

    Ahmad Alamri & Jan K. Blancato

  2. Department of Clinical Laboratories Sciences, College of Applied Medical Sciences, King Khalid University, Abha, Saudi Arabia

    Ahmad Alamri

  3. Daegu Catholic University Medical Center, Daegu, South Korea

    Jun Yeb Nam

  4. Georgetown Lombardi Comprehensive Cancer Center, Fisher Center for Hereditary Cancer and Clinical Genomics Research, Georgetown University, 3300 Whitehaven Street, NW, Suite 4100, Washington, DC, 20007-2401, USA

    Jan K. Blancato

Authors
  1. Ahmad Alamri
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  2. Jun Yeb Nam
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  3. Jan K. Blancato
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Corresponding author

Correspondence to Jan K. Blancato .

Editor information

Editors and Affiliations

  1. Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USA

    Virginia Espina

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Alamri, A., Nam, J.Y., Blancato, J.K. (2017). Fluorescence In Situ Hybridization of Cells, Chromosomes, and Formalin-Fixed Paraffin-Embedded Tissues. In: Espina, V. (eds) Molecular Profiling. Methods in Molecular Biology, vol 1606. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6990-6_17

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  • DOI: https://doi.org/10.1007/978-1-4939-6990-6_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6989-0

  • Online ISBN: 978-1-4939-6990-6

  • eBook Packages: Springer Protocols

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