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Pre- and Postnatal Diagnostics and Research on Peripheral Blood, Chorion, Amniocytes, and Fibroblasts

  • Anja WeiseEmail author
  • Thomas Liehr
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Peripheral blood, amniocytic fluid, chorion, and fibroblasts are the tissues most frequently used for chromosome studies. All four of them are relatively easy to obtain and simply brought into short-term culture, and metaphase spreads can be prepared within a relatively short time. Such metaphase spreads, as well as the huge amounts of previously superfluous interphase nuclei in cytogenetic preparations, are materials that are very well suited to FISH analyses. All kinds of FISH probes can be used to analyze metaphase spreads, while the interphase nuclei can only be analyzed for routine purposes by satellite and locus-specific probes. As the quality and quantity of metaphases is important in research and routine diagnostics, the preparation procedure for the chromosomes is outlined in detail. Moreover, a standard two-color FISH approach, which includes the possibility of enhancing the DAPI-banding pattern by BrdU during cultivation, is described.

Keywords

Chromosome Preparation Metaphase Spread Hypotonic Solution Hypotonic Treatment Inverse Microscope 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

Supported in part by the Friedrich Schiller University Jena, the Ernst-Abbe-Stiftung and Dr. Robert Pfleger-Stiftung.

References

  1. Claussen U, Michel S, Mühlig P, Westermann M, Grummt UW, Kromeyer-Hauschild K, Liehr T (2002) Demystifying chromosome preparation and the implications for the concept of chromosome condensation during mitosis. Cytogenet Genome Res 98:136–146CrossRefPubMedGoogle Scholar
  2. Eisenberg B, Wapner RJ (2002) Clinical procedures in prenatal diagnosis. Best Pract Res Clin Obstet Gynaecol 16:611–627CrossRefPubMedGoogle Scholar
  3. Ford CE, Hamerton JL (1956) The chromosomes in man. Nature 178:1020–1030CrossRefPubMedGoogle Scholar
  4. Gardner RJM, Sutherland GR (2004) Chromosome abnormalities and genetic counseling, 3rd edn. Oxford University Press, New YorkGoogle Scholar
  5. Hliscs R, Mühlig P, Claussen U (1997) The spreading of metaphases is a slow process which leads to a stretching of chromosomes. Cytogenet Cell Genet 76:167–171CrossRefPubMedGoogle Scholar
  6. Hsu TC (1952) Mammalian chromosomes in vitro: 1. karyotype of man. J Heredity 43:167–172Google Scholar
  7. Hughes A (1952) Some effects of abnormal toxicity on dividing cells in chick tissue cultures. Quart J Mikrosc Sci 93:207Google Scholar
  8. Jarrett KL, Michaelis RC, Phelan MC, Vincent VA, Best RG (2001) Microsatellite analysis reveals a high incidence of maternal cell contamination in 46, XX products of conception consisting of villi or a combination of villi and membranous material. Am J Obstet Gynecol 185:198–203CrossRefPubMedGoogle Scholar
  9. Kohne E. (2007) 50 years Kleihauer–Betke test. Klin Padiatr 219:252–253CrossRefPubMedGoogle Scholar
  10. Moorhed PS, Hsu TC (1956) Cytologic studies of HeLa, a strain of human cervical carcinoma. III. Durations and characteristics of the mitotic phases. J Natl Cancer Inst 16:1047–1066Google Scholar
  11. Moorhead PS, Nowell PC, Mellman WJ (1960) Chromosome preparations of leukocytes cultured from human peripheral blood. Exp Cell Res 20:613–616CrossRefPubMedGoogle Scholar
  12. Nowell PC (1960) Phytohemagglutinin: An initiator of mitosis in cultures of normal human leukocytes. Cancer Res 20:462–466PubMedGoogle Scholar
  13. Spurbeck JL, Zinsmeister AR, Meyer KJ, Jatal SM (1996) Dynamics of chromosome spreading. Am J Med Genet 61:387–393CrossRefPubMedGoogle Scholar
  14. Tjio JH, Levan A (1956) The chromosome number of man. Hereditas 42:6–9Google Scholar
  15. Verma RS, Babu A (1994) Human chromosomes: principles and techniques, 2nd edn. McGraw-Hill, New YorkGoogle Scholar
  16. Wegner RD (1999) Diagnostic cytogenetics. Springer, BerlinGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2009

Authors and Affiliations

  1. 1.Institut für Humangenetik und AnthropologieJenaGermany

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