Greetings: I am working up a flow cytometry method for heparin induced thrombocytopenia (HIT), using incubation of normal platelets and heparin in the presence of patient plasma, followed by measurement of platelet activation using CD62p. This is following the method of Tomer (Journal of Hematology, 1997, 98, 648-656). The source article describes production of a positive control by incubation of platelets with calcium ionophore A23817. I am having some difficulty getting this to work, and details in the article are scanty. Does anyone have experience with use of calcium ionophore for this purpose? I'm particularly interested in: - Which salt of CaI is best (a Sigma number would be ideal)? - What is your process for preparation of the CaI (actual recipe)? - How long are platelets incubated with CaI, and are they washed prior to labeling with CD62p? - What are the stability and optimal storage conditions for CaI in solution? Thanks very much for your assistance! Michael Suter, MT Flow Cytometry Tech Specialist Oregon Medical Laboratories msuter@omlabs.com
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:25:57 EST