Figure S1.

Two-Photon Optogenetic Stimulation of Targeted Populations, Non-PC Control Group Characterization, Stimulation Resolution, and Brain Motion Quantification, Related to Figure 1

(A) Regions of interest extracted from one example session FOV. (B) 5 spatially clustered groups of Reward-PCs overlaid on a correlation image of the GCaMP6f signal from the imaging FOV, pixels values are weighted by their signal correlation with neighboring pixels. (C) Stimulation triggered average ΔF/F average intensity image from artifact subtracted recording frames during stimulation, taken from the same session as (A), red circles indicate targeted Reward-PCs, white circles denote non-targeted Start-PCs and blue circles denote stimulation zone place cells. (D) Average response magnitude across the population of responsive target neurons from each session. (E) Percentage of targets deemed responsive from each session. (F) Stimulation specificity for each experimental session, defined as the number of responsive neurons in the group divided by the sum of the number of responsive neurons across all groups. (G) Place cell population average ΔF/F across virtual space for targeted non-place cells and place cells from the same sessions, neurons in both plots are ordered by their peak ΔF/F in odd trials. (H) Event rates of place cells, targeted non-place cells and other non-place cells both in the virtual reality world and during the inter-trial interval, events were defined as the ΔF/F reaching 3 SD above the mean, error bars are SEM. (I) X and Y axis optical point spread function measurement for a stimulation beamlet. (J) Axial optical point spread function measurement for a stimulation beamlet. (K) All-optical physiological resolution along the X/Y axis as measured by the normalized GCaMP6f signal resulting from stimulation at different X/Y displacements from the target neurons, n = 21 neurons from 3 mice, error bars are 95% confidence interval. (L) Axial all-optical physiological resolution as measured by the normalized GCaMP6f signal resulting from stimulation at different Z displacements from the target neurons, n = 23 neurons from 2 mice, error bars are 95% confidence interval. (M) The fraction of off-target neurons per targeted cell when stimulating at different axial displacements, n = 2 sessions from 2 mice. (N) X/Y displacement resulting from brain motion during all stimulation epochs, determined by the translation required to maximize the correlation between each frame and an averaged image of the FOV.