How about inflammatory mediators to activate platelets, such as histamine, thrombin, or phorbol ester. The gene expression of p-selectin is best measured at 10 min (temp dependent). P-selection can be measured in flow (CD62P) , but be careful when optimizing assay because there seems to be a secondary surge of P-selectin after the initial raise declines. Best, Masa Sasagawa, ND Naturopathic Physician - Washington Affiliate Assistant Professor - University of Washington School of Nursing, Dept of Biobehavioral Nursing and Health Systems Postdoctoral Research Fellow - Bastyr University 14500 Juanita Dr NE, #472 Kenmore, WA 98028 Phone: 425-602-3178 Fax: 425-602-3079 masas@bastyr.edu ________________________________ From: Nebe-Von-Caron, G [mailto:g.nebe-von-caron@unipath.com] Sent: Tuesday, November 08, 2005 7:21 AM To: cyto-inbox Subject: RE: [ Platelet activation question ] you can measure platelet activation by their binding to monocytes which is a marker for heart risk, but cell cell aggregation should not be confused with platelet activation as it can occur independently particularly when washing and centrifugation steps are part of the sample preparation. You can measure that separately by a range of surface markers shown on the platelet, but you have to be aware of the problems of sampling and sample storage to prevent the platelets going off. When measuring platelet - leukocyte aggregation one has to distinguish true binding events from coincident events. In case of heart risk the binding to the monocots should be specific, e.g be higher in percentage than for the other cells, easies seen in a log side scatter versus the respective CD markers. Not having researched the field in detail but looked ad adhesion molecules some time ago I would just want to make people aware that adhesion molecule expression is a question of anticoagulant and time and temperature, and so should be cell adhesion. There are a lot of confusing articles out about the phenomenon of cell to platelet aggregation that lack systematic controls. Regards Gerhard -----Original Message----- From: KurtzJ@usa.redcross.org [mailto:KurtzJ@usa.redcross.org] Sent: 07 November 2005 15:17 To: Cytometry Mailing List Subject: [ Platelet activation question ] I would like to know is anyone out there has a procedure to measusare platelet activation using leukocyte aggregates? I also have seen that the measurement of leukocyte aggregates is usually done for invivo studies, is this also an appropriate measurement for invitro work? Thanks, any comments would be appreciated. Jim KurtzReceived on Wed Nov 9 12:38:00 2005
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