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Perceiving mitosis in eukaryotic cells

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Summary

A sensitive method has been developed for visualizing eukaryotic cells in mitosis (M) phase. It employs Zenker's fixative, which makes the plasma membrane but not the nuclear envelope permeable to immunoglobulins. Zenker's-fixed cells are exposed to an antibody which recognizes a major constituent of chromatin. In this case the antibody is a monoclonal (MC 21) which recognizes histone H2b. Because cells in M phase do not have an intact nuclear envelope, the antibody has access to and interacts with their chromatin. The presence of a nuclear envelope in Zenker's-fixed interphase cells precludes access of the antibody to the nuclear chromatin. Consequently, this indirect immunofluorescence procedure selectively labels M-phase cells. At high enough magnification some details of the chromatin figures are revealed. MC 21 recognizes the chromatin of cells of many different species. With appropriate fixation it can be used effectively on cells in culture. With some procedural modifications it can also be used with more complex tissue systems. Detailed mitotic patterns for chick embryos up to Day 3 of development have been obtained by this method.

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Author notes

  1. Dennis Byrne

    Present address: Department of Botany, Iowa State University, Ames, IA

  2. Paul Hwang

    Present address: Johns Hopkins Medical School, Baltimore, MD

Authors and Affiliations

  1. Department of Biochemistry, University of Kansas, 66045, Lawrence, Kansas

    Hae-Young Kim, Dennis Byrne, Paul Hwang, Sandra Collins Thompson & Paul A. Kitos

Authors
  1. Hae-Young Kim
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  2. Dennis Byrne
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  3. Paul Hwang
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  4. Sandra Collins Thompson
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  5. Paul A. Kitos
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Additional information

These studies were supported in part by grant ES 03651 from the National Institutes of Health, Bethesda, MD, and a grant from the Fogarty International Center between the NIH (USA) and CNRS (France).

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Kim, HY., Byrne, D., Hwang, P. et al. Perceiving mitosis in eukaryotic cells. In Vitro Cell Dev Biol 24, 100–107 (1988). https://doi.org/10.1007/BF02623886

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  • DOI: https://doi.org/10.1007/BF02623886

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