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Thrombus Formation In a Living Mouse

Hemostasis and thrombosis are complex processes that involve the vessel wall, circulating blood cells, soluble blood coagulation and regulatory proteins, and blood cell-derived microparticles. Although previous work in this field has employed purified systems to parse this complexity and to allow development of models for the workings of these processes, it has become increasingly important to analyze hemostasis and thrombosis in a living animal where all of the components are present simultaneously. The development of knockout technology in the mouse and improvements in microscopic imaging instrumentation and high speed computers have allowed the development of a system for high speed intravital digital videomicroscopy in the microcirculation of a living mouse. The goal has been to visualize thrombus formation following vessel wall injury in the microcirculation of a living mouse using an intravital confocal and widefield microscopy imaging system. By revisiting and testing the models and hypotheses that have been developed from in vitro studies, we have been able to determine that many of these models and hypotheses are correct whereas some may need some significant modification. The study of biochemistry and cell biology in a living animal offers opportunities to understand physiology and pathology in complex biological systems. Over the years in vitro studies of blood coagulation and vascular biology have taught us what reactions and interactions can happen, but now it is time to learn what does happen in vivo. We have entered a new phase of discovery–an area that is developing rapidly: the ability to study thrombosis in living animals. Historically, our laboratory has employed biochemical, immunochemical and cell biology techniques to parse the mysteries of blood coagulation– with a special focus on the vitamin K-dependent blood coagulation proteins. When we surrendipitously discovered the protein now known as Pselectin [1], our work expanded to include platelets. Upon the discovery that P-selectin is an adhesion molecule that binds platelets to leukocytes [2] and the