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Fig. 7. Migratory behavior of neutrophils in the hi2217;MPO:GFP fish. (A) Single frame from time-lapse movie (Movie 1 in supplementary material) of hi2217;MPO:GFP embryo tailfin at 2 dpf, fluorescence overlaid onto DIC images. Three individual tracks of GFP-positive neutrophils are shown, with the start of each track indicated with an arrow. Dashed boxes indicate selected segments of tracks, depicted in B and C. Bar, 100 µm. (B,C) Time-lapse sequences of tracks from A, with times indicated at the top of each image in hours:minutes:seconds; the side of each image box equals 50 µm. Large arrows indicate the direction of motility, small arrowheads mark pseudopodia. Note how mutant neutrophils frequently stop and extend pseudopodia. (D) Higher magnification (box side, 25 µm) sequence of a single neutrophil; otherwise same as B,C. Note how several pseudopodia are extended over a 2-minute period. (E) Plot of average velocity against percentage of time stopped (percentage of distance data points less than 0.75 µm) for full-length mutant tracks; each point indicates the parameters for an individual cell, dashed lines indicate the average values (in parentheses) for each data set. (F-I) Analysis of hi2217 mutant neutrophil tracks in 15-minute windows. Single frames from time-lapse movies of hi2217;MPO:GFP embryo tailfins at 2 dpf are shown; fluorescence overlaid onto DIC images. Individual tracks of GFP-positive neutrophils are shown, with the start of each track indicated by an arrow; bars, 100 µm. Next to each track image is a plot of directional persistence (D/T, blue lines) and velocity (µm/minute, red lines) over 15-minute time intervals (x axis) for the length of each track. D/T values are multiplied by 10 to use the same numerical y axis as velocity; therefore D/T values above 7 (dashed lines) indicate directional migration.
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