Figure 2.
Figure 2. Platelet, TF, and fibrin deposition in thrombi of wild-type and low TF mice. (A) TF, fibrin, and platelet colocalization in a representative thrombus in a low TF mouse and a wild-type mouse. Upper panels are wild-type mouse; lower panels, low TF mouse. TF was detected with an anti–mouse TF antibody and Alexa 488–conjugated chicken anti–rabbit IgG. An Alexa 660–conjugated rat anti–mouse CD41 and an Alexa 350–conjugated anti–mouse fibrin antibody labeled the platelets and fibrin, respectively. In all experiments, infusion of anti-CD41 was used to detect platelets and antifibrin-specific antibodies were used to detect fibrin. Anti-TF antibodies or irrelevant IgG were infused, as indicated. The thrombus is shown at varying stages of its development: 0%, before vessel wall injury; at 25% of maximal thrombus size; at 50% of maximal thrombus size (early phase); at 100% (peak) thrombus size; and at the plateau phase. Intravital 4-channel video images depict the accumulation of platelets (red), TF (green), and fibrin (blue) as well as the colocalization of TF and platelets (yellow), platelets and fibrin (magenta), TF and fibrin (turquoise), and all 3 components (white). (B) Ratio of the integrated fluorescence intensity associated with tissue factor (FTF) to the integrated fluorescence intensity associated with platelets (FP) in thrombi generated in wild-type and low TF mice. All values are depicted as a percentage of the integrated fluorescence intensity in wild-type mice. Nonspecific background fluorescence was determined using a nonimmune rabbit IgG. (C) Ratio of the integrated fluorescence intensity associated with fibrin (FF) to integrated fluorescence intensity associated with platelets (FP) in thrombi formed in low TF and wild-type mice. The integrated fluorescence intensity in the absence of fibrin formation was determined in a low TF mouse treated with hirudin (2 U/g body weight). All values shown are median values. In panels B and C, 3 mice of each genotype were used in these experiments to generate 19 thrombi in wild-type mice (WT), 18 thrombi in low TF mice (LTF), and 8 thrombi in low TF mice treated with an irrelevant antibody or with hirudin. *P < .05; **P < .025 with respect to wild-type mice. Anti-TF indicates antitissue factor antibodies.

Platelet, TF, and fibrin deposition in thrombi of wild-type and low TF mice. (A) TF, fibrin, and platelet colocalization in a representative thrombus in a low TF mouse and a wild-type mouse. Upper panels are wild-type mouse; lower panels, low TF mouse. TF was detected with an anti–mouse TF antibody and Alexa 488–conjugated chicken anti–rabbit IgG. An Alexa 660–conjugated rat anti–mouse CD41 and an Alexa 350–conjugated anti–mouse fibrin antibody labeled the platelets and fibrin, respectively. In all experiments, infusion of anti-CD41 was used to detect platelets and antifibrin-specific antibodies were used to detect fibrin. Anti-TF antibodies or irrelevant IgG were infused, as indicated. The thrombus is shown at varying stages of its development: 0%, before vessel wall injury; at 25% of maximal thrombus size; at 50% of maximal thrombus size (early phase); at 100% (peak) thrombus size; and at the plateau phase. Intravital 4-channel video images depict the accumulation of platelets (red), TF (green), and fibrin (blue) as well as the colocalization of TF and platelets (yellow), platelets and fibrin (magenta), TF and fibrin (turquoise), and all 3 components (white). (B) Ratio of the integrated fluorescence intensity associated with tissue factor (FTF) to the integrated fluorescence intensity associated with platelets (FP) in thrombi generated in wild-type and low TF mice. All values are depicted as a percentage of the integrated fluorescence intensity in wild-type mice. Nonspecific background fluorescence was determined using a nonimmune rabbit IgG. (C) Ratio of the integrated fluorescence intensity associated with fibrin (FF) to integrated fluorescence intensity associated with platelets (FP) in thrombi formed in low TF and wild-type mice. The integrated fluorescence intensity in the absence of fibrin formation was determined in a low TF mouse treated with hirudin (2 U/g body weight). All values shown are median values. In panels B and C, 3 mice of each genotype were used in these experiments to generate 19 thrombi in wild-type mice (WT), 18 thrombi in low TF mice (LTF), and 8 thrombi in low TF mice treated with an irrelevant antibody or with hirudin. *P < .05; **P < .025 with respect to wild-type mice. Anti-TF indicates antitissue factor antibodies.

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