Abstract
FISH (fluorescent in situ hybridization) is a molecular cytogenetic technique established in the early 1980s that allows for the detection of DNA copy number changes (gains and losses) mapping to genomic regions of interest (Langer-Safer et al. Proc Natl Acad Sci USA 79:4381–4385, 1982). This technology has been extensively applied to research-based investigations and is routinely used in prenatal diagnosis and oncology. Here we describe a modification of the standard FISH protocol adapted for the detection of low-frequency mosaic aneuploidy in interphase cells. This approach represents a straightforward method for the measurement of aneuploidy levels in mammalian cells. This system combines four probes mapping to two different chromosomes. The choice of probes is essential for the successful performance of this approach. It greatly reduces the enumeration of false-positive signals that are challenging in the enumeration of ploidy changes (particularly if these are complex and/or involve a significant increase of chromosome number).
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References
Langer-Safer PR, Levine M, Ward DC (1982) Immunological method for mapping genes on Drosophila polytene chromosomes. Proc Natl Acad Sci U S A 79:4381–4385
Hudziak RM, Lewis GD, Winget M, Fendly BM, Shepard HM et al (1989) p185HER2 monoclonal antibody has antiproliferative effects in vitro and sensitizes human breast tumor cells to tumor necrosis factor. Mol Cell Biol 9:1165–1172
Slamon DJ, Leyland-Jones B, Shak S, Fuchs H, Paton V et al (2001) Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. N Engl J Med 344:783–792
Faggioli F, Wang T, Vijg J, Montagna C (2012) Chromosome-specific accumulation of aneuploidy in the aging mouse brain. Hum Mol Genet 21:5246–5253
Faggioli F, Vezzoni P, Montagna C (2011) Single-cell analysis of ploidy and centrosomes underscores the peculiarity of normal hepatocytes. PLoS One 6:e26080
Duncan AW, Taylor MH, Hickey RD, Hanlon Newell AE, Lenzi ML et al (2010) The ploidy conveyor of mature hepatocytes as a source of genetic variation. Nature 467:707–710
Rehen SK, McConnell MJ, Kaushal D, Kingsbury MA, Yang AH et al (2001) Chromosomal variation in neurons of the developing and adult mammalian nervous system. Proc Natl Acad Sci U S A 98:13361–13366
Westra JW, Peterson SE, Yung YC, Mutoh T, Barral S et al (2008) Aneuploid mosaicism in the developing and adult cerebellar cortex. J Comp Neurol 507:1944–1951
Freshney RI (2010) Culture of animal cells: a manual of basic technique and specialized applications. 6th ed. New York: Wiley-Liss; 187–206
Kent WJ, Sugnet CW, Furey TS, Roskin KM, Pringle TH et al (2002) The human genome browser at UCSC. Genome Res 12: 996–1006
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, Vols 1, 2 and 3
ASI ASI GenASIs Capture & Analysis. http://www.spectral-imaging.com/platforms/genasis-capture-a-analysis
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Faggioli, F., Vijg, J., Montagna, C. (2014). Four-Color FISH for the Detection of Low-Level Aneuploidy in Interphase Cells. In: Sharp, D. (eds) Mitosis. Methods in Molecular Biology, vol 1136. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0329-0_14
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DOI: https://doi.org/10.1007/978-1-4939-0329-0_14
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0328-3
Online ISBN: 978-1-4939-0329-0
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