Paolo- I've been using Molecular Probes Live/dead kits for about 2yrs now with great success. Although I use #4 (runs off the UV) I've also tried #2 which works well off 488nm. You incubate the cells for 30min, then are able to fix to analyze later. Good Luck! bunny Bunny Cotleur, M.S. Sr Research Technologist, Dept of Neurosciences Scientific Consultant, Flow Cytometry Core Cleveland Clinic Foundation Lerner Research Institute, NC30 9500 Euclid Avenue Cleveland, OH 44195 Lab: (216)444-1164 ********************************************* Caminante, no hay camino. Se hace camino al andar. On Oct 21, 2005, at 8:41 AM, paolo wrote: > Dear Fellow Flowers, > > I work with HIV infected cultures and I need to determine apoptotic > vs live vs dead cells. I use an Coulter XL with a single laser (488nm) > and I have to fix cells with 1% PFA as they are loaded with HIV. If I > use PI or 7AAD all cells will turn dead after fixation. Can anyone > suggest alternatives to these dyes? I already tried EMA, but perhaps > there something else out there that does not require the extra step > with a light source... > > Thanks in advance for all your help. > -- > > ============================ > Aut viam inveniam aut faciam > ============================ > Paolo Piazza, PhD > OFFICE: 604 Parran Hall > tel (412) 383 - 9590 > LABORATORY: 529 Parran Hall > tel: (412) 648 - 2940 > Infectious Diseases and Microbiology > GSPH – University of Pittsburgh > 130 Desoto Street > 15261 Pittsburgh PA > fax (412) 624 – 4953 > e-mail paolo@pitt.edu > > >Received on Mon Oct 24 15:18:00 2005
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