Intravital confocal microscopy reveals impaired platelet–neutrophil interactions in anti-GPVI–treated mice after LPS treatment. (A) Automated tracking of platelets interacting with neutrophils with Imaris Bitplane software. Example images derived from videos of the time interval from 25 to 30 minutes, showing probability masks of neutrophils superimposed with platelet fluorescent signal and platelet tracks (left), or platelet fluorescence signal superimposed with platelet tracks (middle), or platelet tracks only (right) of the indicated group. Blue tracks, firm interacting platelet; and red tracks, short interacting platelets. Scale bar, 5 μm. (B-C) Fraction of interacting platelets that showed firm interaction (B) and fraction of total platelet tracks that showed firm interaction with neutrophils and are thus considered as PNC (C). Each data point represents 1 mouse at the respective time interval. Bars represent mean time interval ± standard deviation. The two-way ANOVA with a Tukey multiple comparisons test. ∗P ≤ .05; ∗∗P < .01.