Barbara, I don't know much about lectins and rbcs, but we have run some cultured cells stained with a FITC conjugated lectin, and there was a definite dose-dependent toxic effect. If the lectin concetration is high you may just be killing all the cells, which would account for decreased scatter (especially FSC). Our staining with this lectin (don't know which one) was intense--1000x brighter than the negatives, so they were able to use concentrations that didn't kill to high a percentage. BTW, this was the only time I have had to worry about FITC overlap into the red channel (>610). We used PI for viability, and it took me a while to realize that our FITC labelled cells were showing up as dimly "red", even w/o PI! Just my few cents, Steve --------------------------------------------------------------- Steve G. Hilliard steve@habanero.cb.uga.edu If you're a runner you should be dead. http://storm.cadcam.iupui.edu/drs/drs.html --------------------------------------------------------------- On Mon, 24 Nov 1997, Barbara Butcher wrote: > > happy thanksgiving! > > a colleague is looking at the ability of rbcs from various species to bind > fluorescent lectins by facs. when she adds some lectins to the rbcs, the > cells lose forward and side scatter. i would expect fsc and ssc to > increase, if the cells are agglutinating. any suggestions? > > thanks! > > barbara > > ****************************************************************************** > Barbara A. Butcher, Ph.D. > James A. Baker Institute for Animal Health > Hungerford Hill Road > Cornell University > College of Veterinary Medicine > Ithaca, New York 14853 > > (607) 256-5647 Lab > (607) 256-5618 Office > (607) 256-5608 FAX > **************************************************************************** > ** > >
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