UNIT 7.24 Detection of Mitotic Cells
Published Online: 1 MAY 2004
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2004
- 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;
- multiparameter laser scanning cytometery;
- mitotic index