Supplementary Figure 7: Simultaneous mesoscale volumetric functional imaging of distant neurites and synapses.

(a) Maximum intensity projection of a Gaussian image stack of neurites in an awake mouse cortex, with features color-coded by depth. FOVs P1 and P2 (white squares) were separated by ~ 2mm and selected for additional random-access functional imaging. (b) (Left panels) Maximum intensity projections of Gaussian image stacks at P1 and P2, with each volume extending over 400 µm × 425 µm × 200 µm and composed of 101 images with 0.5 µm pixel size and a z step of 2 µm. Features color-coded by depth. Volume rate: 0.1 Hz. (Right panels) Zoomed-in views of the structures within the four colored squares. Post-objective power: 36-48 mW, increased exponentially with depth. (c) Composite of 2 images acquired by scanning a Bessel focus at 0.5 µm per pixel and a z step size of 100 µm. Volume rate: 5.1 Hz. (Right panels) Zoomed-in views of the structures within the four colored squares. Post-objective power: 120 mW. (d) Raster plots of calcium transients detected from all spines and boutons in P1 and P2.