Extended Data Fig. 1: The spatiotemporal property of microglia motility after FLA injury.
a. Illustration of experiments in which microglia motility was monitored in awake Cx3cr1GFP/+ mice in vivo. Focal laser ablation (FLA) was used to introduce local injury, and two-photon imaging was conducted to track microglia. b. Representative images of microglia prior to FLA (−FLA) or after FLA at indicated time points. c-e. Time-dependent changes of microglia migration after FLA. The migration index is quantified as the relative fluorescence change in the injury center (180 μm in diameter) over the whole image. The data of each mouse are shown in (c), and averaged data as well as the changes of migration index over time (Δmigration index) are plotted in (d) (n = 5 mice). The persistence is the time with effective signal over baseline in (e) (n = 5 mice). f. Illustration of the measurement of microglia polarity. g. The changes of microglia polarity to FLA at the single cell level. h. Illustration of the microglia directionality towards the injury center. The angle between the microglia polarity and injury is quantified. i. The microglia directionality before (−FLA) and after FLA (n = 366 microglia from 5 mice for −FLA; n = 313 microglia from 5 mice for +FLA). j. The relationship between microglia directionality and the distance of microglia to injury. The area with signal above baseline (−FLA) is defined as coverage and quantified (n = 5 mice). Scale bars, 50 µm. Data are shown as the mean ± s.e.m. ***, p < 0.001; All statistical tests are two-sided and see Supplementary Table 1 for statistics.
