Figure 3. The effect of anesthesia on microglia surveillance differs with pathology and age in V1.

a Representative in vivo two-photon images showing fewer microglial pixels in time projected images in awake animals (reflecting lower microglial surveillance) than in anesthetized animals at 4 months (left) and to a lesser extent at 9 months (right) (scale bar = 25μm). Plaque-associated microglia (outlined in yellow) were manually selected based on proximity to MeX04⁺ plaques (not shown). b Representative in vivo two-photon images showing the selection of plaque-associated microglia (outlined in yellow). c-e Quantification of microglial surveillance in awake versus anesthetized state in CX3CR1^GFP/+ (Control) and CX3CR1^GFP/+ 5xFAD^+/− (5xFAD) mice (c: 4 months; d: 6 months; e: 9 months). f Representative images of manually selected individual microglia from awake and anesthetized mice for Sholl analysis. g Representative Sholl curves showing Sholl profiles of microglia in awake (black) and anesthetized (green) control mice at 4 months. h-j Both control and plaque-distal microglia, but not plaque-associated microglia, are more ramified (more total intersections) under anesthesia (h: 4 months; i: 6 months; j: 9 months). n = 8–10 mice per genotype per age group. Repeated measures ANOVA with Bonferroni (c-e) or Tukey (h-j) correction; *p<0.05, **p<0.01, ***p<0.001.