Figure 3.
Figure 3. Neutrophils and monocytes are the main leukocyte subsets accumulating during the initiation of DVT. (A) Neutrophils (green) crawling on the vessel wall (red) of the IVC 2 h after DVT induction visualized by two-photon microscopy. Tracks of individual neutrophils are shown in white (also see Video 3). Bar, 50 µm. Shown is a representative of n = 3 experiments. (B) Time course of early leukocyte–endothelial interaction within 6 h of flow restriction as evaluated by intravital microscopy in WT animals using Acridine orange (WT + acr orange). Baseline images were taken before ligation. LysM-eGFP mice were used to evaluate neutrophils and CX3CR1-eGFP mice were analyzed to evaluate monocytes. Bars, 100 µm. Shown is a representative of n = 5 experiments. (C) Dynamics of the recruitment of distinct subsets of innate immune cells during DVT initiation determined in vivo by video microscopy. Rolling and firm adhesion of leukocytes are given as number per square millimeter. Results are shown as mean ± SEM (n = 5 per group). (D, Left) Relative frequency of neutrophils and monocytes at 6 h of flow restriction as assessed by intravital video microscopy. Results are mean ± SEM (n = 5 per group). (D, Right) Representative images taken by intravital epifluorescence microscopy showing blood cell recruitment 6 h after flow restriction in the IVC. Neutrophils were visualized in LysM-eGFP mice. The number of recruited monocytes was assessed using CX3CR1-eGFP mice. In both strains of mice, all leukocytes (irrespective of their lineage) were identified by counterstaining with the fluorescent dye Acridine orange (pseudocolored in red). Bars, 100 µm. (E) FACS analysis of blood in LysM-eGFP and CX3CR1-eGFP mice without IVC ligation using a neutrophil specific anti-Ly6G antibody. (F) Intravital two-photon microscopy of LysM-eGFP and CX3CR1-eGFP with PE-labeled anti-Ly6G antibody 6 h after DVT induction (left). Bars, 50 µm. Quantification of Ly6G+ GFP+ double-positive cells in these mouse strains (right; Videos 4 and 5). Shown is a representative of n = 3 experiments.

Neutrophils and monocytes are the main leukocyte subsets accumulating during the initiation of DVT. (A) Neutrophils (green) crawling on the vessel wall (red) of the IVC 2 h after DVT induction visualized by two-photon microscopy. Tracks of individual neutrophils are shown in white (also see Video 3). Bar, 50 µm. Shown is a representative of n = 3 experiments. (B) Time course of early leukocyte–endothelial interaction within 6 h of flow restriction as evaluated by intravital microscopy in WT animals using Acridine orange (WT + acr orange). Baseline images were taken before ligation. LysM-eGFP mice were used to evaluate neutrophils and CX3CR1-eGFP mice were analyzed to evaluate monocytes. Bars, 100 µm. Shown is a representative of n = 5 experiments. (C) Dynamics of the recruitment of distinct subsets of innate immune cells during DVT initiation determined in vivo by video microscopy. Rolling and firm adhesion of leukocytes are given as number per square millimeter. Results are shown as mean ± SEM (n = 5 per group). (D, Left) Relative frequency of neutrophils and monocytes at 6 h of flow restriction as assessed by intravital video microscopy. Results are mean ± SEM (n = 5 per group). (D, Right) Representative images taken by intravital epifluorescence microscopy showing blood cell recruitment 6 h after flow restriction in the IVC. Neutrophils were visualized in LysM-eGFP mice. The number of recruited monocytes was assessed using CX3CR1-eGFP mice. In both strains of mice, all leukocytes (irrespective of their lineage) were identified by counterstaining with the fluorescent dye Acridine orange (pseudocolored in red). Bars, 100 µm. (E) FACS analysis of blood in LysM-eGFP and CX3CR1-eGFP mice without IVC ligation using a neutrophil specific anti-Ly6G antibody. (F) Intravital two-photon microscopy of LysM-eGFP and CX3CR1-eGFP with PE-labeled anti-Ly6G antibody 6 h after DVT induction (left). Bars, 50 µm. Quantification of Ly6G+ GFP+ double-positive cells in these mouse strains (right; Videos 4 and 5). Shown is a representative of n = 3 experiments.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal