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Histochemistry

, Volume 74, Issue 1, pp 57–61 | Cite as

A reliable preparation of mono-dispersed chromosome suspensions for flow cytometry

  • M. Stoehr
  • K. J. Hutter
  • M. Frank
  • Kl. Goerttler
Article

Summary

A reproducible, simple and rapid protocol was developed to prepare mono-dispersed chromosome suspensions for flow cytometric analysis. The procedure basically employes: 1. twice rinsing of monolayer cultures to eliminate dead cells and cellular debris, 2. mild fixation of mitotic cells with 1% acetic acid, and 3. ultra sonic treatment to release single metaphase chromosomes. The procedure takes less than 30 min. Isolated chromosomes are storable over months at 4° C. Despite mild fixation many biochemical studies and experiments applied on such fixed chromosomes purified and enriched using electronic sorting are feasible: 1. gene and restriction mapping, 2. cloning of specific gene sequences, and 3. gene frequency analysis.

Keywords

Acetic Acid Dead Cell Frequency Analysis Flow Cytometric Analysis Gene Frequency 
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.

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References

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Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • M. Stoehr
    • 1
  • K. J. Hutter
    • 1
  • M. Frank
    • 1
  • Kl. Goerttler
    • 1
  1. 1.Institute for Experimental PathologyGerman Cancer Research Center HeidelbergHeidelbergGermany

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