Detection of collagen type I and tissue factor in thrombi after FeCl3 or laser-induced injury. (A) Injuries in wild-type or FcRγ-null mice were induced by application of 10% FeCl3 for 5 minutes or by laser. Platelets were labeled by injection of Alexa 660–conjugated rat anti-CD41 antibody into the circulation prior to injury (red pseudocolor). Collagen was labeled by injection of a rabbit anti–mouse collagen type I antibody (1 μg/g body weight) plus an Alexa 488–conjugated goat anti–rabbit antibody (1 μg/g body weight) prior to injury (green pseudocolor). Merge of platelets and collagen, yellow pseudocolor. (B-C) Cremaster muscles injured by FeCl3 or by laser were isolated and sectioned. Image sections of a thrombus were acquired at 0.5-μm intervals across the vessel diameter perpendicular to the long axis of the vessel by incrementally altering the working distance between the microscope objective and the speciment with a piezo-electric driver.23 (B) Tissue sections were stained with Alexa 660 anti–mouse CD41 antibody, rabbit anti–mouse collagen type I antibody, and an Alexa 488–conjugated goat antirabbit antibody. Platelets are indicated by red pseudocolor; collagen, green pseudocolor; and the merge, yellow pseudocolor. Pictures shown are representative of 4 independent experiments. (C) Tissue sections were stained with Alexa 660–conjugated anti–mouse CD41 antibody, rabbit anti–mouse tissue factor (TF) antibody, and Alexa 488–conjugated goat antirabbit antibody. Platelets are indicated by red pseudocolor; tissue factor, green pseudocolor; and the merge, yellow pseudocolor. Pictures shown are representative of 4 independent experiments.
