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

Lyons, A. Bruce, Blake, Stephen J. and Doherty, Kathleen V. (2013) Flow cytometric analysis of cell division by dilution of CFSE related dyes. Current Protocols in Cytometry, Supp.64: 9.11.1-9.11.12. doi:10.1002/0471142956.cy0911s64

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Author Lyons, A. Bruce
Blake, Stephen J.
Doherty, Kathleen V.
Title Flow cytometric analysis of cell division by dilution of CFSE related dyes
Journal name Current Protocols in Cytometry   Check publisher's open access policy
ISSN 1934-9297
1934-9300
Publication date 2013-04
Year available 2013
Sub-type Article (original research)
DOI 10.1002/0471142956.cy0911s64
Open Access Status
Issue Supp.64
Start page 9.11.1
End page 9.11.12
Total pages 12
Place of publication Hoboken, NJ, United States
Publisher Jossey Bass
Collection year 2014
Language eng
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.
Keyword Adoptive transfer
Cell division
CFSE
CTV
Flow cytometry
Proliferation
Q-Index Code CX
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Lab protocols.

Document type: Journal Article
Sub-type: Article (original research)
Collections: Non HERDC
ERA White List Items
UQ Diamantina Institute Publications
 
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