From: Len Brown (lenbrown@all.com.au)
Date: Tue Feb 10 1998 - 23:21:02 EST
Dear Ray and friends Don't know why the method in Ormerod did not work, were the samples refrigerated at any time? Otherwise, why can't you use a whole blood method? The use of whole blood will eliminate most causes of platelet aggregation and agglutination. See: Michelson,AD Flow cytometry: A clinical test of platelet function. Blood 87,12:4925 Len Brown Ray Hicks wrote: > > Hi, > > I've been asked to post this message to the list, has anyone got any > reliable preps for platelets? Any ideas with what might have gone wrong > with the cited method? > > TIA > > Ray > > >A method for preparing isolated platelets for FACS analysis (we will be > >looking to see if a particular protein is expressed on the surface of these > >cells). > > > >We will be using blood from humans and from pigs in relatively small > >volumes (tens of mls rather than litres; individual samples not pooled) and > >have access to cold rooms, centrifuges, etc. etc. > >We followed the method given in 'Flow Cytometry - A Practical Approach' by > >Ormerod 2nd edition; but this gave us un-resuspendable platelet pellets and > >relatively high red blood cell contamination. > > > >If anyone has a reliable prep method, or knows where we are likely to have > >gone wrong in following the textbook method mentioned we will be very > >grateful. > > Ray Hicks > ________________________________________________________________________ > |University of Cambridge |Tel 01223 330149 | > |Department of Medicine |Fax 01223 336846 | > |Level 5, Addenbrookes Hospital |e-mail <rh208@cus.cam.ac.uk> | > |Hills Road Cambridge |Web http://facsmac.med.cam.ac.uk | > |CB2 |ftp server ftp://131.111.80.78 | > |UK | | > |_________________________________|_____________________________________|
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