I've had luck with Sigma's PKH67-GL though it took some amount of time for me to determine that the package insert's range is toxic. I seem to recall that a 40-fold dilution did the trick without cellular toxicity and with sufficient labeling to be bright. William C. William King, Jr. Technical Coordinator Flow Cytometry FACS core research facility Children's Research Institute, Room 5220 Children's National Medical Center 111 Michigan Ave., NW Washington, District of Columbia, 20010 202-884-2308 202-884-5808 fax wking@cnmc.org <<< "Reed, Doug S Dr USAMRIID" <Doug.Reed@DET.AMEDD.ARMY.MIL> 7/10 4:17p >>> PKH-26 is also fairly toxic to cells, at least in our hands. That's why we use CFSE instead. -Doug Douglas S. Reed, Ph.D. Microbiologist Respiratory & Mucosal Immunity Department of Aerobiology & Product Evaluation Division of Toxinology & Aerobiology U.S. Army Medical Research Institute of Infectious Diseases 1425 Porter Street, Fort Detrick Frederick, MD 21702-5011 301-619-6728 301-619-6911 fax doug.reed@det.amedd.army.mil -----Original Message----- From: Joanne Lannigan [mailto:joannelannigan@virginia.edu] Sent: Monday, July 08, 2002 10:15 AM To: cyto-inbox Subject: RE: tracking cell division, not CFSE Hi Simon: You could try PKH-26 a lipophilic dye which excites at 488 and emits in the red/orange range. It is often used for cell tracking /proliferation assays. I believe it is sold by Sigma. Regards, Joanne Joanne Lannigan, MS Director, Flow Cytometry Core Facility Jordan Hall, Room 7067 P.O. Box 800734 Charlottesville, VA 22908-0734 Office: 434-924-0274 Lab: 434-263-2695 Fax: 434-982-1071 email: joannelannigan@virginia.edu > -----Original Message----- > From: Simon Monard [mailto:smonard@trudeauinstitute.org] > Sent: Tuesday, July 02, 2002 2:31 PM > To: Cytometry Mailing List > Subject: tracking cell division, not CFSE > > > Hi folks > > Has anyone had any success using both CFSE and another dye to track cell > division of > two ex-vivo labelled cell populations.We use CFSE all the time and it > works great, > what we need is another orange or red fluorescing dye that like CFSE will > be retained > by cells in vivo and will allow us to follow the number of cell > divisions. We have > tried Snarf but it seems to be toxic to the cells we are using. The other > dye must > be excitable with either 488nm or 633nm. Any suggestions? > > best wishes > > Simon > > Simon Monard > FACS Lab Manager > Trudeau Institute > Saranac Lake > NY12983 > > Ph 518 891 3080 X352
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