BACKGROUND: Plateletpheresis components have been shown to contain p-selectin-positive platelets after collection and storage. P-selectin mediates binding of activated platelets to granulocytes and monocytes. This study was undertaken to assess platelet activation, granulocyte activation, platelet-granulocyte heterotypic aggregate formation, and the plasma-soluble p-selectin level during plateletpheresis performed on a particular instrument (MCS+, Haemonetics). STUDY DESIGN AND METHODS: Flow cytometry was used to assay platelet surface p-selectin, granulocyte iC3b receptor, and platelet-granulocyte aggregates in the platelet component, residual blood in the disposable polycarbonate bowl of the MCS+, and in the donor blood with and without the addition of in vitro agonists before, during, and after plateletpheresis. The plasma-soluble p-selectin levels in the platelet component, disposable bowl, and donor venous blood were measured by an enzyme-linked immunosorbent assay. RESULTS: Levels of p-selectin-positive platelets, activated granulocytes, and platelet-granulocyte aggregates were greater in the disposable bowl than in the preapheresis donor blood. Levels of p-selectin-positive platelets, activated granulocytes, and platelet-granulocyte aggregates in the postapheresis donor blood were similar to those in the preapheresis donor blood. The platelet components contained no activated granulocytes or detectable platelet-granulocyte heterotypic aggregates, and only about 10-percent activated platelets. The plasma-soluble p-selectin level in the platelet component was significantly greater than that in the preapheresis donor blood, the residual blood in the disposable bowl, or the postapheresis donor blood. CONCLUSIONS: Measurements of platelet surface p-selectin, platelet-granulocyte heterotypic aggregates, and plasma-soluble p-selectin can be used to detect platelet activation during plateletpheresis.