Dear Flowers and Flowerettes, I have a question for those of you with expertise on CFSE: I am setting up CFSE staining of sheep lymphocytes for LTT and have trouble in obtaining a decent UNIFORM staining of my cells. In other words, the CV of each peak (population) in FL1 is very wide (included the unstimulated control-single peak). I had tested the same protocol in the past (8 months ago) and at that time I had quite good results. The only difference between then and now is that my CFSE stained cells are resuspended in a different media AFTER the CFSE staining (IMDM instead of RPMI). Can the change in growth media influence the amount of CFSE retained by my cells? Or is it more likely that I'm setting my FACS wrong? or something odd in my reagents? I load CFSE in HBSS using pluronic, CFSE final concentration is between 2.5 an 5 micromolar, 20' of loading. Any help will be greatly appreciated, and I'll post a summary of the answers for the list as usual. Thank you very much for your help, Mara *********************** Dr. Mara S.L. Rocchi Moredun Research Institute International Research Centre Pentland Science Park Bush Loan Penicuik EH26 0PZ Scotland UK e-mail: roccm@mf.mri.sari.ac.uk Tel: 0131-4455111 ext. 464/461 Fax: 0131-446235 "The information contained in this email is confidential. It may also be legally privileged. It is intended only for the stated addressee(s) and access to it by any other person is unauthorised. If you are not an addressee, you must not disclose, copy, circulate or in any other way use or rely on the information contained in this email. Such unauthorised use may be unlawful. If you have received this email in error please reply to it immediately and delete it and all copies from your system."
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:57:30 EST