Reply to Walter Sharp

T. Vincent Shankey tshanke at bsd.meddean.luc.edu
Fri Mar 1 15:39:07 EST 1996


Walter,

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 
further there. 
Vince Shankey



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