CFSE

From: Julie Oughton (oughtonj@ucs.orst.edu)
Date: Tue Oct 13 1998 - 16:30:25 EST

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Good afternoon!

I really could use some help with a new application. I have an investigator
who first stains 10 million mouse lymphocytes with CFSE (carboxyfluorescein
diacetate succinimidyl ester dissolved in DMSO; final concentration = 5 ug
CFSE per ml; 0.1% DMSO) in a total volume of 1 ml, washes the cells in
PBS/10% FBS, and then stains for cell surface expression of CD4. 

It appears that we have a number of problems. First of all, treatment with
CFSE drastically alters the light scatter measurements and CD4 expression
appears to be lost. Secondly, CSFE staining is so bright that we have a
difficult time compensating FL1 fluorescence out of FL2.

Does any one have any experience with CFSE staining? Any suggestions?

Thanks in advance,
Julie Oughton
Oregon State University

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