gfp and facs analysis

Gerstein, Rachel Rachel.Gerstein at umassmed.edu
Tue Oct 12 17:04:16 EST 1999


The best way is to analyze live cells that are not fixed, in the presence of
1 ug/ml propidium iodide to exclude dead cells. Dont use a hard gate to
exclude PI+ cells - instead look at FL3 vs FL plots to gate on GFP+,
PI-negative.
=======================================================
Rachel M. Gerstein, Ph.D.
Department of Molecular Genetics and Microbiology
Graduate Program in Immunology/Virology
University of Massachusetts Medical School
55 Lake Avenue North
Worcester, MA 01655-0002
(508) 856-1044
(508) 856-5920 (FAX)


> ----------
> From: 	Nicholas Sibinga
> Sent: 	Tuesday, October 12, 1999 12:57 PM
> To: 	Cytometry Mailing List
> Subject: 	gfp and facs analysis
> 
> 
> Hello -- 
> 
> I'm hoping that you can give me some advice or protocol information on
> FACS analysis of transiently-transfected cells expressing green
> fluorescent protein. I am planning to use a co-transfected EGFP plasmid to
> serve as a marker for transfection, but I am concerned that fixation
> methods (such as those used for propidium iodide analysis of DNA content)
> may decrease the GFP signal. Can you advise the best way to do this? 
> 
> Thanks very much, 
> 
> Nicholas Sibinga, MD 
> 
> Cardiovascular Biology Lab 
> Harvard School of Public Health 
> 677 Huntington Avenue 
> Boston, MA 02115 
> 
> phone 617-432-3586 
> fax 617-432-2980 
> 
> 



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