We have used confocal and widefield microscopy to image thrombus formation in real time in the microcirculation of a living mouse. This system provides high-speed, near-simultaneous acquisition of images of multiple fluorescent probes and of a brightfield channel. Vascular injury is induced with a laser focused through the microscope optics. We observed platelet deposition, tissue factor accumulation and fibrin generation after laser-induced endothelial injury in a single developing thrombus. The initiation of blood coagulation in vivo entailed the initial accumulation of tissue factor on the upstream and thrombus–vessel wall interface of the developing thrombus. Subsequently tissue factor was associated with the interior of the thrombus. Tissue factor was biologically active, and was associated with fibrin generation within the thrombus.
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We thank S. Inoué for introducing us to the Yokogawa CSU-10 high-speed confocal microscope, Eric Furie for digital image capture advice, and F. Castellino and E. Rosen for demonstrating the laser injury model. This work was supported by grants from the US National Institutes of Health (HL51926 and HL69435). The confocal microscope was obtained with partial support from the National Institutes of Health (S10RR15680).
The authors declare no competing financial interests.
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Falati, S., Gross, P., Merrill-Skoloff, G. et al. Real-time in vivo imaging of platelets, tissue factor and fibrin during arterial thrombus formation in the mouse. Nat Med 8, 1175–1180 (2002) doi:10.1038/nm782
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