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In Situ Hybridization for DNA: Fluorescent Probe

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Book cover Molecular Histochemical Techniques

Part of the book series: Springer Lab Manuals ((SLM))

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Abstract

Fluorescence in situ hybridization (FISH) with chromosomespecific probes is one of the non-isotopic in situ hybridization methods that opens up several new areas of cytogenetic investigation by allowing visual determination of the presence and normality of specific genetic sequences in single metaphase or interphase cells (Pinkel 1986a, 1986b). With this approach, the genetic loci to be analyzed are made microscopically visible in single cells using in situ hybridization with nucleic acid probes specific to these loci. The DNA in target cells is made as a single strand and incubated with single-stranded fluoro chromedirectly- labeled or chemically modified DNA probe under conditions where the probe will anneal only with DNA sequences to which it has high DNA sequence homology. The bound chemically modified probe is made visible by immunofluorescent detection methods. This technique is often called standard FISH.

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References

  • Du Manoir S, Kallioniemi O.P., Lichter P et al (1995) Hardware and software requirements for quantative analysis of comparative genomic hybridization. Cytometry 19:4–9

    Article  PubMed  Google Scholar 

  • Houldsworth J, Chaganti R.S.K. (1994) Comparative genomic hybridization: An overview. Am J Pathol145:1253–1260

    PubMed  CAS  Google Scholar 

  • Kallioniemi A, Kallioniemi O.P., Sudar D, Rutovits D, Gray J.W., Waldman F.M., Pinkel D (1992) Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors. Science 258:818–821

    Article  PubMed  CAS  Google Scholar 

  • Kallioniemi O.P., Kallioniemi A, Kurisu W, Thor A, Yu L.C., Smith H.S., Waldman F.M., Pinkel D, Gray J.W. (1992) ERBB2 amplification in breast cancer analyzed by fluorescence in situ hybridization. Proc. Natl. Acad. Sci. USA. 89; 5321–5325

    Article  PubMed  CAS  Google Scholar 

  • Kallioniemi A, Kallioniemi O-P, Sudar D, Rutovits D, Gray J.W., Waldman F.M., Pinkel D (1993) Comparative genomic hybridization: A rapid new method for detecting and mapping DNA amplification in tumors. Semin Cancer Biol4:41–46

    PubMed  CAS  Google Scholar 

  • Kallioniemi O.P., Kallioniemi A, Piper J, Isola J, Waldma, F.M., Gray J.W., Pinkel D (1994) Optimizing comparative genomic hybridization for analysis of DNA sequence copy number changes in solid tumors. Gene Chrom Cancer 10: 231–243

    Article  CAS  Google Scholar 

  • Matsuta M, Matsuta M, Hayashi S, Yasumi S, Sasaki K, Kuroda Y, Nishiya I (1996) Reconstructed Three-dimensional Images of Flow-sorted Nuclei Hybridized with Chromosome-specific Probes. J Histochm Cytochem 44: 1337–1343

    Article  CAS  Google Scholar 

  • Matsuta M, Matsuta M, Kon S, Thompson C, LeBoit P.E., Weier H.U., Gray J.W. (1994) Interphase cytogenetics of melanocytic neoplasmic numerical aberrations of chromosomes can be detected in interphase nuclei using antromeric DNA probes. J Cutan Pathol 21:1–6

    Article  PubMed  CAS  Google Scholar 

  • Persons D.L., Hartmann L.C., Herath J.F., et al. (1993) Interphase molecular cytogenetic analysis of epithelial ovarian carcinomas. Am J Path 142:733–741

    PubMed  CAS  Google Scholar 

  • Pinkel D, Gray J.W., Trask B, van den Engh G, Fucoe J, van Dekken, H (1986a) Cytogenetic analysis by in situ hybridization with fluorocently labeled nucleic acid probes. Cold Spring Harbor Symp Quant Biol 51: 151–157

    Article  PubMed  CAS  Google Scholar 

  • Pinkel D, Straume T, Gray J.W. (1986b) Cytogenetic analysis using quantitative, high sensitivity fluorescence in situ hybridization. Proc Natl Acad Sci USA 83: 2934–2938.

    Article  PubMed  CAS  Google Scholar 

  • Piper J, Rutovits D, Sudar D, et al (1995) Computer image analysis of comparative genomic hybridization. Cytometry 9:10–26

    Article  Google Scholar 

  • Qian J, Bostwick D.G., Takahashi S, et al.(1995) Chromosomal anomalies in prostatic intraepithelial neoplasia and carcinoma detected by fluorescence in situ hybridization. Cancer Res 55:5408–5414

    PubMed  CAS  Google Scholar 

  • Schroeck E, Veldman T, Padilla-Nash H et al. (1997) Spectral karyotyping refines cytogenetic diagnostics of constitutional chromosomal abnormalities. Human Genetics 101:255–262

    Article  Google Scholar 

  • White D.M., Crolla J.A., Ross F.M. (1995) Detection of minimal residual disease in childhood acute lymphoblastic leukemia using fluorescence in situ hybridization. Br J Haematol 91:1019–1024

    Article  PubMed  CAS  Google Scholar 

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© 2000 Springer Japan

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Matsuta, M., Matsuta, M. (2000). In Situ Hybridization for DNA: Fluorescent Probe. In: Koji, T. (eds) Molecular Histochemical Techniques. Springer Lab Manuals. Springer, Tokyo. https://doi.org/10.1007/978-4-431-67915-8_6

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  • DOI: https://doi.org/10.1007/978-4-431-67915-8_6

  • Publisher Name: Springer, Tokyo

  • Print ISBN: 978-4-431-68520-3

  • Online ISBN: 978-4-431-67915-8

  • eBook Packages: Springer Book Archive

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