Re: rbc and lectin study by facs

From: Steve G. Hilliard (steve@habanero.cb.uga.edu)
Date: Mon Nov 24 1997 - 19:45:03 EST


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
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