Laser-induced cremaster arteriole thrombosis model

Male mice (8-10 weeks of age) were anesthetized by intraperitoneal injection of ketamine/xylazine (100 mg/kg), and a tracheal tube was inserted to facilitate breathing.^10,17 A jugular vein catheter was established to inject antibodies and reagents. Anti-platelet (DyLight 488 anti-GPIb; 1 μg/g) and anti-fibrin (Alexa Fluor 647; 0.3 μg/g) antibodies were administered prior to intravital microscopy. Compounds were administered for 10 minutes prior to induction of vascular injury using a laser ablation system (Ablate! photoablation system; 3i). Images were acquired in real-time, using a 63× water-immersion objective, with a Zeiss Axio Examiner Z1 multichannel fluorescent microscope equipped with a solid laser launch system and high-speed sCMOS camera. Images were analyzed using SlideBook software.