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. 2017:1606:265-279.
doi: 10.1007/978-1-4939-6990-6_17.

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

Ahmad Alamri  1   2 , Jun Yeb Nam  3 , Jan K Blancato  4   5
Affiliations

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

Ahmad Alamri et al. Methods Mol Biol. 2017.

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.

Keywords: DNA probes; Fluorescence microscopy; Formalin-fixed paraffin-embedded tissues; Metaphase chromosomes.

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Figures

Figure 1
Figure 1
Step by step FISH experiment. The target DNA and fluorochrome-labeled DNA probe are denatured and allowed to hybridize. The slide is then washed, counterstained, and viewed with fluorescent microscopy
Figure 2
Figure 2
Fluorescent in situ hybridization images. (a) Breast cancer tissue hybridized with a control chromosome 8 centromeric probe labeled in green and a MYC probe labeled in red showing low-level amplification. DAPI counterstain in blue. (b) Normal prostate cell metaphase chromosomes labeled with red and green unique sequence DNA probes. DAPI counterstain in blue
See this image and copyright information in PMC

References

    1. Kallioniemi A, Visakorpi T, Karhu R, Pinkel D, Kallioniemi OP. Gene copy number analysis by fluorescence in situ hybridization and comparative genomic hybridization. Methods. 1996;9(1):113–121. - PubMed
    1. Pinkel D, Gray JW, Trask B, van den Engh G, Fuscoe J, van Dekken H. Cytogenetic analysis by in situ hybridization with fluorescently labeled nucleic acid probes. Cold Spring Harb Symp Quant Biol. 1986;51(Pt 1):151–157. - PubMed
    1. Waye JS, Willard HF. Chromosome-specific alpha satellite DNA: nucleotide sequence analysis of the 2.0 kilobasepair repeat from the human X chromosome. Nucleic Acids Res. 1985;13(8):2731–2743. - PMC - PubMed
    1. Cajulis RS, Frias-Hidvegi D, Yu GH, Eggena S. Detection of numerical chromosomal abnormalities by fluorescence in situ hybridization of interphase cell nuclei with chromosome-specific probes on archival cytologic samples. Diagn Cytopathol. 1996;14(2):178–181. doi: 10.1002/(SICI)1097-0339(199603)14:2<178::AID--DC14>3.0.CO;2-J. - DOI - PubMed
    1. Cremer T, Lichter P, Borden J, Ward DC, Manuelidis L. Detection of chromosome aberrations in metaphase and interphase tumor cells by in situ hybridization using chromosome-specific library probes. Hum Genet. 1988;80(3):235–246. - PubMed

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