Reply to Walter Sharp
T. Vincent Shankey
tshanke at bsd.meddean.luc.edu
Fri Mar 1 15:39:07 EST 1996
I'm still digesting the issue you raise concerning mean fluorescence
intensity in thrombocytopoienia. My main concern there is how will you
standardize the fluorescence signal "per platelet".
Meanwhile, regarding the issue of large list mode files... We take
advantage of an old time flow trick (you have to be over 40 to use this
one)- we stain the whole blood preperation with CD61 or CD41a labelled
with FITC and acquire the data with a threshold descriminator on the FITC
channel set to include only labelled platelets (works on our Epics minus
5 and on the XL). This way the list mode file is not occupied by 90% RBC
or WBC (unless they have a pletelet stuck to them- you must take care
here to prove that you've diluted your sample sufficiently to rule out
coincidental counting of particles). Note- we do not use a FALS
descriminator. Also, if your particles have a mutated GPIIb/IIIa that is
not recognized at all by your monoclonal Ab, you have a serious problem.
I would be happy to stop by and discuss this at further length with
you... but I don't get over to your neck of the "woods" much these days.
If you are going to the ISAC meeting at Rimini, I'd be happy to talk
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