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Prinzip der DNS-Cytometrie zur Bestimmung des nucleären DNS-Gehalts von Zellkernsuspensionen

Anwendungsmöglichkeiten mittels Ethidiumbromid/Mithramycin-Fluorochromierung

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Neue Entwicklungen in der Dermatologie

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Zusammenfassung

Abweichungen des cellulären DNS-Gehalts (DNS-Aneuploidie) sind, zusammen mit hohen S-Phase- und Mitoseraten, wichtige Kennzeichen neoplastischer Gewebe. Die Bestimmung dieser Parameter mit Hilfe der DNS-Cytometrie hat sich in der onkologischen Forschung durchgesetzt. Das Verfahren erlaubt die schnelle und präzise Messung des nucleären DNS-Gehalts von isolierten Zellen in Suspension.

In der vorliegenden Arbeit werden Prinzipien der Durchflußcytometrie dargestellt, wobei auf die stöchiometrische DNS-Fluorochromierung mit Ethidiumbromid und Mithramycin und deren quantitative Messung mit dem Durchflußcytophotometer ICP 22 näher eingegangen wird.

Fehlermöglichkeiten bei der DNS-Analyse sowie deren Vermeidung und Behebung werden, ebenso wie weitere Anwendungsmöglichkeiten der Durchflußcytometrie, kurz besprochen.

Summary

DNA-aneuploidy and both raised S-phase and mitotic rates are proven to be important characteristics of human neoplasms. Due to velocity and high accuracy, quantitative DNA measurements by aid of flow systems hold an important place in oncologic research. This technique allows the rapid and accurate determination of the DNA amount in large single cell suspensions.

The present paper, focussing on the stoichiometric DNA fluorochromation with Ethidiumbromide and Mithramycin and the quantitative DNA measurements in the flow cytometer ICP 22, outlines principles of Flow Cytometry.

Technical problems and pitfalls of DNS analysis are pointed out and discussed together with additional applications of Flow Cytometry.

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Authors and Affiliations

  1. Dermatologische Klinik mit Poliklinik, Universität Erlangen-Nürnberg, Hartmannstraße 14, D-8520, Erlangen, Deutschland

    T. Arnold & E. Deinlein

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  1. T. Arnold
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  2. E. Deinlein
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  1. Erlangen, Deutschland

    O. P. Hornstein

  2. Münster-Hornheide, Deutschland

    M. Hundeiker

  3. Mannheim, Deutschland

    J. Schönfeld

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Arnold, T., Deinlein, E. (1988). Prinzip der DNS-Cytometrie zur Bestimmung des nucleären DNS-Gehalts von Zellkernsuspensionen. In: Hornstein, O.P., Hundeiker, M., Schönfeld, J. (eds) Neue Entwicklungen in der Dermatologie. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-73581-3_4

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  • DOI: https://doi.org/10.1007/978-3-642-73581-3_4

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