Figure 3
From: A new approach for ratiometric in vivo calcium imaging of microglia
Post hoc analyses of activation markers expressed in Twitch-2B labelled microglia. (a,b) MIP images of fixed cortical tissue, transduced in vivo with LV.CMV.Twitch-2B.miR-9.T viral vector, taken in the epicentre (a; 7–59 µm, step 1 µm) and at the periphery (b; 4–44 µm, step 1 µm) of the injection site. Left: Twitch-2B positive microglial cells labelled with an anti-GFP antibody. Middle: the same field of view labelled with anti-CD68 antibody. Merged images are shown on the right. (c,d) MIP images of fixed cortical tissue (as above) taken in the epicentre (c; 2–28 µm, step 1 µm) and at the periphery (d; 1–33 µm, step 1 µm) of the injection site. Left: Twitch-2B positive microglial cells labelled with an anti-GFP antibody. Middle: the same field of view labelled with anti-IL-1β antibody. Merged images are shown on the right. Inset in (d) shows a rare Twitch-2B and IL-1β positive cell at the periphery of the injection site. (e) Summary box plot illustrating distributions of background-subtracted CD68 fluorescence per microglial cell in the epicentre (n = 100 cells in 3 mice) and at the periphery (n = 86 cells in 3 mice) of the injection site. Note that CD68 fluorescence is significantly higher in microglia in the epicentre compared to periphery (p < 0.001, Mann-Whitney test). (f) Summary bar graph showing the fraction of IL-1β positive cells in the population of Twitch-2B positive microglia in the epicentre (n = 31 cells in 4 mice) and at the periphery (n = 54 cells in 5 mice) of the injection site.
