Pre- and Postnatal Diagnostics and Research on Peripheral Blood, Chorion, Amniocytes, and Fibroblasts
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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.
KeywordsChromosome Preparation Metaphase Spread Hypotonic Solution Hypotonic Treatment Inverse Microscope
Supported in part by the Friedrich Schiller University Jena, the Ernst-Abbe-Stiftung and Dr. Robert Pfleger-Stiftung.
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