Stability and stationarity of spontaneous GCaMP6 activity in functional connectivity analysis.

(A) Functional connectivity maps were constructed using time windows of spontaneous 0.4–4.0Hz GCaMP6 (awake) data of increasing length from one mouse (Mouse 2). Windows were increased in length by ~2.6s up to ~300s. Spatial similarity (Dice coefficient) was calculated between each of these functional connectivity maps for all networks relative to their corresponding group level map. The functional connectivity maps calculated by using a seed in left motor cortex are highlighted as an example. Across all networks, similarity scores converge at 0.9 with a window length of ~30s. (B) Seed-based functional connectivity maps from Mouse 2 constructed using 30s of spontaneous GCaMP6 data. (C) For each of 7 networks, the same 30s window of spontaneous data for each network was compared to the corresponding group-level network functional connectivity map for each of seven mice (circles). The stability of similarity of 30s of data is present across network and across mice. (D) Using consecutive 30s windows (sampled every ~2.6s) from a single 300s imaging session from each mouse, the spatial similarity between all 30s motor functional connectivity maps and the group level motor functional connectivity map is shown to be relatively stationary. Error bars are standard error of the mean.