Distribution of adhesive proteins and platelets in thrombi generated in perfusion of normal control blood under a low or high shear rate condition.
Whole blood from a healthy control containing mepacrine-labeled platelets was perfused over a collagen surface with a low (300 s−1) or high (1500 s−1) shear rate for 7 minutes. Fixed thrombi on coverslips were stained with a Cy3-labeled anti-VWF or antifibrinogen antibody and analyzed by CLSM. Merged CLSM images were obtained by the superimposition of 2 images at the identical portion and slice of thrombi; green indicates platelets in thrombi; orange indicates VWF (left panels) and fibrinogen (right panels). CLSM images displayed in rows 1, 2, and 3 are cross-sections at a height of 30, 15, and 0 μm from the collagen surface, respectively (original magnifications, × 400). Images are representative of 5 independent perfusions using blood from 5 individual donors. Under high shear rate conditions, VWF is predominantly distributed at the outer areas of thrombi, whereas fibrinogen is present at the inner areas. In contrast, only fibrinogen appears in thrombi under low shear rate conditions.
