Unit

UNIT 9.11 Flow Cytometric Analysis of Cell Division by Dye Dilution

  1. A. Bruce Lyons1,
  2. Kathleen V. Doherty2

Published Online: 1 FEB 2004

DOI: 10.1002/0471142956.cy0911s27

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Lyons, A. B. and Doherty, K. V. 2004. Flow Cytometric Analysis of Cell Division by Dye Dilution. Current Protocols in Cytometry. 27:9.11:9.11.1–9.11.10.

Author Information

  1. 1

    Institute of Medical and Veterinary Science, Adelaide, South Australia

  2. 2

    Australian Red Cross Blood Service, Adelaide, South Australia

Publication History

  1. Published Online: 1 FEB 2004
  2. Published Print: JAN 2004

This is not the most recent version of the article. View current version (1 APR 2013)

Abstract

The technique described in this unit uses the intracellular fluorescent label carboxyfluorescein diacetate succinimidyl ester (CFSE) to tag proliferating cells. Covalently bound CFSE is divided equally between daughter cells, allowing discrimination of successive rounds of cell division. The technique is applicable to in vitro cell division as well as to in vivo division of adoptively transferred cells and can resolve up to eight successive generations. CFSE is long lived, permitting analysis for several months after transfer, and has the same spectral characteristics as fluorescein, so monoclonal antibodies conjugated to phycoerythrin or other compatible fluorochromes may be used to immunophenotype the dividing cells.

Keywords:

  • flow cytometry;
  • cell division;
  • CFSE;
  • proliferation;
  • adoptive transfer