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1) whole blood technique for Platelet surface markers where red cell lysis
is performed post marking ?
2) Glanzmann's thrombasthenia - Anyone using flow for detection of
heterozygous
states ? "Correction" of Mean Florescence Index to "equalise" patients and
normals
using FS or MPV appears to hold some promise.
To: cyto-inbox
1) We fix whole blood in 1% PFA, then wash in Tyrodes-Hepes buffer prior to
labeling platelets. This buffer tends to promote gentle RBC lysis at 4oC. We
have some data regarding the commercial lyse-fix reagents in platelet
labeling. We found that platelet fixation is critical if samples are to be
stored or if there is a lysis step at any point other than simply using the
Tyrodes buffer. I'd recommend fixing the sample before lysis; whether you
need to fix before marking depends on your storage time and antibodies.
2) We have seen only homozygous Glanzmann's. Sounds like a great project.
From: henry.rinder@yale.edu
Ph. 203-785-4231
Fx. 203-737-4111
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web