Procoagulant activity of BDMPs from freeze-thawing injury. (A) (Upper) TEM image of BDMPs (bar = 200 nm). (Inset) A microparticle stained with an immune-gold-labeled GFAP antibody (bar = 100 nm). (Lower) TEM image of BDMPs with negative staining to enhance the view of membrane structures (bar = 200 nm). (B) BDMPs and platelet microparticles solubilized in an SDS lysis buffer were probed for NSE, GFAP, and von Willebrand factor (T, whole brain lysate; Pts, platelet lysate as control). (C) The expression of TF, PS, and both was measured for BDMPs (represents 20 separate measurements). (D) PS-dependent clotting time was measured in the presence of increasing numbers of BDMPs (n = 6, 1-way ANOVA, P < .001) and compared with those induced by 1.6 μg/μL of purified brain PS and PC, respectively. (E) Plasma clotting times induced by 25 000/μL BDMPs were measured in the presence of increasing concentrations of bovine lactadherin. (F) TF-dependent thrombin generation was measured in reaction containing either 1 pM TF or 25 000 BDMPs (representative of 3 separate experiments).