Multi-parameter assessment of thrombus formation on microspotted arrays of thrombogenic surfaces

Abstract

Abstract Thrombus formation by adhering and aggregating blood platelets is fundamental to hemostasis and is a prerequisite for vascular occlusion in pathological thrombosis. The parallel-plate flow chamber technique has been extensively used to measure platelet adhesion and activation in vitro at arterial or venous flow conditions. Here, we describe the use of brightfield and confocal fluorescence microscopy to record the various platelet activation processes contributing to thrombus formation on microspotted arrays of thrombogenic surfaces; and we give procedures to analyze the acquired microscopic images. Furthermore, we describe technical problems that can be expected using the microspot technique. Content: (A) Flow chamber preparation and whole blood perfusion. (B) Brightfield and fluorescence microscopic imaging of thrombi. (C) Analysis of brightfield and fluorescence images. Introduction Thrombus formation by adhering and aggregating blood platelets is fundamental to hemostasis and is a prerequisite for vascular occlusion in pathological thrombosis.