I am using CFSE and 7-AAD in human lymphocyte cultures without any problems. I can do decent compensation after 3 day cultures. Titrating 7-aad was tedious, as I ended up needing very small conc.(0.15ul/ml). With PI I have no experience in combo with CFSE. Angulo R, Fulcher DA, Cytometry. 1998;34:143-151 is a reference I use for CFSE comparison with thymidine. They did not use a viability dye though. Sanders K. Chai, MD, MPH, FAAP Seattle Biomedical Research Institute 307 Westlake Avenue N, Suite 500 Seattle, WA 98109-5219 USA Telephone: +1 206 256-7200 Fascimile: +1 206 256-7229 Muheza Designated District Hospital c/o Theonest Mutabingwa, MD Private Bag Hospital Road Muheza, Tanga Tanzania Telephone: +255 272 641 420 Mobile: +255 745 483 950 -----Original Message----- From: Henry H. Wortis [mailto:henry.wortis@tufts.edu] Sent: Thu 6/9/2005 1:58 PM To: Cytometry Mailing List Cc: Subject: CFSE, PI, compensation, autofluorescence Can someone lead me to an article or share personal experiences regarding experiments in which CFSE is used together with propidium iodide to study viability and proliferation of lymphocytes over several days in culture? Are there compensation problems? After 3 days in medium (with phenol red) is there a shift in FL3? Under these conditions do CFSE+ cells show a shift in FL3 and therefore appear non-viable? Thanks for your help. henry -- Henry H. Wortis, M.D. Professor & Chair Department of Pathology Director Graduate Program in Immunology Tufts University School of Medicine 150 Harrison Avenue Boston MA 02111 617-636-6718 617-636-2990 (FAX)Received on Mon Jun 13 12:38:00 2005
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