Perceiving Mitosis in Eukaryotic Cells

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 con...

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Veröffentlicht in:In Vitro Cellular & Developmental Biology 1988-02, Vol.24 (2), p.100-107
Hauptverfasser: Kim, Hae-Young, Dennis Byrne, Paul Hwang, Sandra Collins Thompson, Kitos, Paul A.
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Sprache:eng
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Zusammenfassung: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.
ISSN:0883-8364
2327-431X
1475-2689
DOI:10.1007/bf02623886