Perceiving mitosis in eukaryotic cells

Kim Hae-Young Kim, D. Byrne, P. Hwang, S. Collins Thompson, P. A. Kitos

Research output: Contribution to journalArticlepeer-review

Abstract

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 more complex tissue systems. Detailed mitotic patterns for chick embryos up to Day 3 of development have been obtained by this method.

Original languageEnglish (US)
Pages (from-to)100-107
Number of pages8
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume24
Issue number2
DOIs
StatePublished - 1988

ASJC Scopus subject areas

  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

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