Which other fluorochromes do you intend to use in conjunction with CFSE (CFDA SE)? If they are FL3 and/or FL4, compensation might not be a big issue. You will likely have to use some of your CFSE-stained cells (unstained for any other marker) to do your FL1 only compensation (against leakage into FL2). Then you can use cells from an animal that did not receive adoptively transfered cells (or non CFSE-labeled donor cells, whatever these might be) to use in the other singly stained compensation controls (FL2 only, FL3 only etc). Not a perfect solution, but workable. What type of cells and how many did you transfer? The extend of dilution of CFSE (i.e., proliferation) depends entirely upon what cells you are transferring (T cells?, other) and into what host environment (Irradiated, RAG2KO, etc) and how long after transfer you plan on harvesting the cells. I'd look at a publication that performed a similar transfer to get a rough idea of what you might expect in terms of CFSE profiles when you finally do the analysis. I hope this is of some help to you. Larry Wolfraim -----Original Message----- From: Jeffrey Rice To: cyto-inbox Sent: 5/25/2004 9:58 AM Subject: CFDA SE compensation control I have a quick question regarding CFDA SE tracking. I am going to be labeling cells for tracking following adoptive transfer, followed by FACS analysis. How do I deal with the compensation issue, as I will probably not have a significant CFDA SE-positive population in my post-transfer sample, nor will I know the intensity of the staining until I'm on the machine? Jeff -- Jeffrey Rice <rice@aecom.yu.edu> Graduate Student Diamond Lab Albert Einstein College of Medicine, Bronx, NY 10461 v. 718-430-4112 f. 718-430-8711Received on Wed May 26 13:58:00 2004
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