Application of Cryo-High Resolution (Hr) Sem Technique for Imaging Platelet Morphology

Abstract Platelets play a major role in hemostasis due to their ability to adhere and aggregate at sites of vessel wall injury. Damage to the inner surface of arteries includes ruptures or tears at the site of atherosclerotic plaques or injury resulting from interventional procedures. Uncontrolled platelet recruitment and aggregation at these sites can lead to complete occlusion of the vessel. Thus the understanding of platelet function and structure has become an important goal in preventing vascular disease. We employed cryo-HRSEM techniques to gain more insight into the surface morphology of both quiescent and thrombin stimulated platelets. Platelet rich plasma (PRP) was obtained by mixing whole blood (21 ml) drawn from juvenile pigs with 3 ml of acid citrate dextrose and centrifugation at 300g for 10 min. To obtain washed platelets the PRP was washed three times with a citrate wash buffer consisting of 36mM citric acid, 5mM glucose, 5mM KCl, 2 mM CaCl2, 1 mM MgCl2, 90 mM NaCl, pH 6.5.