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Flow cytometric analysis of cell division by dilution of CFSE and related dyes

Citation

Lyons, AB and Blake, SJ and Doherty, KV, Flow cytometric analysis of cell division by dilution of CFSE and related dyes, Current Protocols in Cytometry, John Wiley & Sons, JP Robinson et al (ed), Hoboken, NJ, pp. 9.11.1-9.11.12. ISBN 9780471142959 (2013) [Research Book Chapter]

Copyright Statement

Copyright 2013 Wiley

DOI: doi:10.1002/0471142956.cy0911s64

Abstract

The technique described in this unit uses the intracellular fluorescent label carboxyfluorescein diacetate succinimidyl ester (CFSE) to track 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 eight or more successive generations. CFSE is long lived, permitting analysis for several months after cell 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. In addition, information is given on a second-generation dye, Cell Trace Violet (CTV), excited by 405-nm blue laser light. CTV is chemically related to CFSE, but allows the 488-nm line of the Argon laser to be used for other probes.

Item Details

Item Type:Research Book Chapter
Keywords:flow cytometry, dye dilution, cell division, CFSE, CTV,  proliferation,  adoptive transfer
Research Division:Medical and Health Sciences
Research Group:Immunology
Research Field:Cellular Immunology
Objective Division:Health
Objective Group:Other Health
Objective Field:Health not elsewhere classified
Author:Lyons, AB (Dr Bruce Lyons)
Author:Doherty, KV (Dr Kathleen Doherty)
ID Code:88558
Year Published:2013
Deposited By:Medicine (Discipline)
Deposited On:2014-02-06
Last Modified:2014-12-05
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