UNIT 7.24 Detection of Mitotic Cells

  1. Gloria Juan1,
  2. Zbigniew Darzynkiewicz2

Published Online: 1 MAY 2004

DOI: 10.1002/0471142956.cy0724s28

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Juan, G. and Darzynkiewicz, Z. 2004. Detection of Mitotic Cells. Current Protocols in Cytometry. 28:7.24:7.24.1–7.24.7.

Author Information

  1. 1

    Memorial Sloan-Kettering Cancer Center, New York, New York

  2. 2

    Brander Cancer Research Institute, New York Medical College, Valhalla, New York

Publication History

  1. Published Online: 1 MAY 2004
  2. Published Print: APR 2004


In preparation for cell division, nuclear chromatin undergoes a vital rearrangement required for the organization of chromosomes and their allocation to daughter cells. This process is initiated during G2 phase with the most remarkable morphological manifestation being chromatin condensation. This unit provides protocols for identification and quantification of mitotic cells based on immunocytochemical detection of histone H3 phosphorylated on Ser 10 (H3-P), the critical event occurring during the G2 to M transition (essential for chromatin condensation), using anti-H3-P, a commercially available antibody to which apoptotic cells are not reactive, concurrently with differential staining of cellular DNA. Additionally an adaptation of this method used to stain cells mounted on microscope slides for analysis by multiparameter laser scanning cytometry is also presented.


  • cell cycle;
  • chromatin;
  • histone;
  • multiparameter laser scanning cytometery;
  • mitosis;
  • mitotic index