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Adhesion of platelets to a transparent wall can be observed and recorded continuously by using a shear flow chamber built into a microscope slide. A field of area 50,000-100,000 εm2 can be viewed by phase contrast. Adhesion (and subsequent detachment and departure) events can be monitored as functions of site position in the field and of elapsed time. Fresh anticoagulated human platelet-rich plasma (PRP) was pumped steadily through a flow chamber so that a wall shear rate of 25 sec-1 was obtained at the field of view. Videotapes were analysed for stationary platelets, with the locations of sites and their state of occupancy being digitized for computer data logging and analysis. As the PRP was pumped steadily through the chamber, it was found that there was a 2-3 min delay before adhesion events occurred. Then the cumulative number of adhesion sites grew linearly with time, the least-squares fit giving the cumulative site growth rate (CSGR) with a high correlation coefficient r&gt; 0.98. Some platelets detached, but later the same sites were re-occupied; this process often repeated, so that a few sites were occupied by as many as eight different adherent platelets in fifteen minutes. An index was also found for the re-use of sites (RUS), based on the frequency of re-use. Pre-incubation of platelet-rich plasma 40-105 minutes with aspirin, 5-10 mM, reduced CSGR and RUS. Injection of adenosine, resulting in 50 µM in the flow, and of dipyridamole, 250 µM, both reduced CSGR. Chlorpro- mazine, 10 µM, did not cause significant differences from controls. Preincubation with an analog of cIMP reduced CSGR and RUS markedly, especially in the presence of ADP.

Adhesion Of Human Platelets In Steady Shear Flow: Effects Of Drugs