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