Direct imaging of FVa and platelets. (A) Digital composite fluorescence and brightfield images of representative thrombi in HB mice infused with Alexa₄₈₈-FV-810^SYA (600 µg/kg) at 10, 40, 70, 100, 130, and 190 seconds after injury; Alexa₄₈₈-FV-810^SYA (green), platelets (red), and overlap between FV-810^SYA and platelets (yellow). Bar represents 10 µm. (B-C) Kinetics of Alexa₄₈₈-FV-810^SYA (n = 4 mice; 20 thrombi) and platelets (n = 4 mice; 20 thrombi) fluorescence accumulation over time. (D) Confocal imaging of Alexa₄₈₈-FV-810^SYA and platelets 4 minutes after laser injury, merged or presented individually; step size = 1 µm in z-plane; total section 30 µm. Images depict different orientations of the same thrombus. (D) Upper panel: horizontal orientation; lower panel: 45 degrees rotated. Bar represents 30 µm. Platelet accumulation was monitored using Alexa₅₅₅-labeled rat anti- CD41 F_(ab)2 antibody.