UNIT 9.11 Flow Cytometric Analysis of Cell Division by Dye Dilution
Published Online: 1 FEB 2004
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 FEB 2004
- Published Print: JAN 2004
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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.
- flow cytometry;
- cell division;
- adoptive transfer