We routinely employ the fix-then-stain procedure for human platelets with excellent results, however we have had the same problem that you describe with fixing murine platelets. You must remember that there is a lot of P-Selectin inside the platelet - if your fixation manages to permeabilize the platelet even slightly, you can get apparent CD62P "expression". We have been unable to solve this problem for murine platelets and thus we stain-then-fix for mouse work. If anyone comes up with a successful fixation method for muirne platelets I'd love to hear about it! Ken Ault
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