Automated Microdensitometry And Protein Assays As Measures For Platelet Adhesion And Aggregation On Collagen Coated Slides Under Controlled Flow Conditions
VIIth International Congress on Thrombosis and Haemostasis
Microscope slides were homogeneously coated over a length of 2 cm with a mixture of soluble and fibrillar collagen and exposed at 37°C and under laminar flow to citrated whole rabbit blood at a flow-rate of 100 ml/min. Surface coverage with platelets (adhesion) and platelet accumulations higher than about 5 μm in height (aggregation) were determined by automated microdensitometry of fuchsine stained 'en face' preparations. The platelet mass per unit surface was measured with a modified Lowry
... a modified Lowry technique whose sensitivity was equivalent to 5×l05platelets. Platelet number, amount of protein and surface coverage with platelet accumulations correlated. After a perfusion time of 10 min thrombi up to 30 μm in height and oriented in the direction of flow had developed on the collagen coated area. Surface coverage with platelets was 75% and the amount of deposited protein 1.4 μg/mm2(2×l06platelets/mm2). On the uncoated surface single platelets predominated; the surface coverage was 20% and the density of platelets 8×104/mm2. Acetyl- salicylic acid at 100 μm decreased platelet aggregation by about 80% without affecting adhesion.The new parallel plate perfusion system offers rapid quantitation of platelet-surface and platelet-platelet interaction after exposure to flowing blood and iftay also be diagnostically useful.