Neutrophil migration responses to gradients in mice are similar to those in zebrafish. In support of Fig. 1. (A) Scheme showing two-photon LW chemotaxis assay in mice. Neutrophils from bone marrow of C57Bl6 mice were isolated, fluorescently dye-labeled, and then injected into the ear dermis of C57Bl6-Albino mice. 3 h later, intravital imaging started by recording neutrophil dynamics before and after induction of a small tissue lesion in the skin dermis by a two-photon laser beam. (B) Time-lapse sequence of two-photon confocal image projections, showing CMTPX dye-labeled neutrophils (white) in a C57B16-Albino mouse, migrating pre- and post-LW (red dashed line). LW occurs at 0 min. Scale bar = 50 μm. (C) Cell speed in relation to the cosine of θ for pre- and post-LW. n = 629–4,879 cell steps per bin for pre-wound, n = 2,152–6,673 cell steps per bin for post-wound. (D) Cell cosine of θ in relation to the distance from the closest point of the WP. n = 1,145–3,156 cell steps per bin for pre-wound, n = 1,580–12,000 cell steps per bin for post-wound. (E) Scatter plot of cell δ in relation to θ, color-coded for the density of points, for pre- (top) and post-LW (bottom). Black dashed lines indicate the groups of data points taken for analysis for F. Plotted data from two representative mice. (F) Percentage of cell persistent steps (δ < 45°), small turns (45° < δ < 90°), and U-turns (δ > 90°), respectively, with θ < 60°. Wilcoxon matched-pairs signed rank test, *, P = 0.0156 (for both persistent steps and small teurns). (C, D, and F) Data from seven mice.