Figure 5.

SLS Allows Single AP Resolution during Population Imaging

(A) GCaMP6s-expressing layer IV principal neurons. Yellow circles identify 155 neurons. (A₁) SLS trajectory (yellow line) crossing all the identified cells.

(B and B₁) Fluorescence over time from the two cells indicated in red in (A) in conventional raster scanning acquisitions (B) and SLS (B₁). Blue ticks above traces indicate whisker stimuli (duration: 200 ms).

(C and C₁) Fluorescence over time of all cells shown in (A) (gray traces) during consecutive whisker stimulations and the average across cells (black trace) in raster scanning (C) and SLS (C₁).

(D) SNR of events in all raster scan series and all SLS acquisitions. n = 360 cells from 4 animals; paired t test, p = 5E-4.

(E) Fraction of cells responding to the whisker stimulation in raster scanning and SLS. n = 360 cells from 4 animals; paired t test, p = 4E-12.

(F–F₃) Simultaneous juxtasomal recording and raster scan experiment from a GCaMP6s-expressing neuron at 10 Hz (F) and 30 Hz (F₁). The white line indicates the recording pipette. The same cell in (F) is recorded using juxtasomal recording and SLS imaging in a much larger FOV at 30 Hz (F₂) and 62.5 Hz (F₃). The yellow line in (F₂) and (F₃) represents the SLS trajectory across the different cells (white number). The same cells were differently numbered in SLS at 30 Hz and 62.5 Hz because the SLS trajectory changed when the surround region was modified to change the scanning rate.

(G) Left: fluorescence over time acquired in raster scanning at 10 Hz (top) and electrophysiology recording (bottom). Right: fluorescence over time acquired in raster scanning at 30 Hz (top) and electrophysiology recording (bottom).

(G₁) The same as in (G) for SLS at 30 Hz (left) and at 62.5 Hz (right).

(H) SNR of fluorescence events associated to isolated APs (see STAR Methods) under the different conditions. n = 16 acquisitions from 8 FOVs in 4 animals; Wilcoxon rank-sum test; p = 5E-4 for raster scan at 10 Hz versus raster scan at 30 Hz; p = 0.12 for raster scan at 30 Hz versus SLS at 30 Hz; p = 7E-3 for SLS at 30 Hz versus SLS at 62.5 Hz.

(I) Accuracy of single AP detection under the different conditions. n = 16 acquisitions from 8 FOVs in 4 animals; Wilcoxon rank-sum test; p = 2E-3 for raster scan at 10 Hz versus raster scanning at 30 Hz; p = 0.01 for raster scan at 30 Hz versus SLS at 30 Hz; p = 2E-3 for SLS at 30 Hz versus SLS at 62.5 Hz.