Hello everyone.
I am having a similar problem, whether I use lectin or antibody. My
problem has been that if I add antibody at low titre, there is no
fluorescence, but if I add it at high titre I find the cells have broken
to pieces. The temperature during labelling has made no difference with
the antibody. I only tried the lectin according to the concentration
suggested by the manufacturer. The rbcs agglutinated and broke apart. I
tried to improve my rbc storage method prior to the labelling to lessen
fragility, but this has not changed results.
Does anyone have a suggestion for fixing the cells prior to labelling? I
have recently tried a method suggested by a fellow subscriber involving
methanol at -70C, however the cells didn't survive my labelling procedure.
I am going to attempt this method once more anyhow. Why are red cells so
fragile??
Thanks.
^..^
_||__(oo)____||___ Monica Nass
-||--"--"----||--- Research Associate
_||_( __ )___||___ Department of Surgery
-||--"--"----||--- Dalhousie University
|| || Halifax, Nova Scotia
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