Figure 3.

Motion Artifacts in SLS

(A) X and Y displacement during raster scanning in anesthetized animals (n = 3 mice, gray), quiet wakefulness (n = 3 mice, orange), and active wakefulness (n = 3 mice, magenta).

(B) Top: representative SLS acquisition during active wakefulness. Pseudocolor scale indicates fluorescence amplitude. Middle: autoregressive second order fit (AR(2), red) of SLS PC1 (black). Bottom: cross correlation (green) between AR(2) and the PC1. The gray arrowhead indicates a movement artifact detected when the 0.3 threshold is crossed.

(C) Representative SLS acquisition with no detected large motion artifacts during quite wakefulness.

(D) Fluorescence over time without (no reassignment, gray) or with (reassigned, red) the reassignment of 11 pixels.

(E) SNR as a function of the number of reassigned pixels for one representative ROI. The gray and the red asterisks indicate the conditions displayed in (D).

(F) Correlation between the motion displacement calculated using NoRMCorre on the reference patch and the motion displacement computed from the pixels reassignment approach (black dots). Empty circles: individual experiments; filled circles: average ± SD. Data from the reference patch were downsampled in time before applying NoRMCorre. Motion displacement values were separately calculated for the X (left) and the Y (right) direction in the reference patch and in the ROIs. The correlation between the motion displacement calculated using NoRMCorre and a random pixels reassignment are shown in gray. Left: downsampling 0.5 Hz, p = 0.08; downsampling 1 Hz, p = 0.18; downsampling 2 Hz, p = 0.06; downsampling 5 Hz, p = 0.32; paired t test, n = 13 SLS acquisitions. Right: downsampling 0.5, p = 2E-6; downsampling 1 Hz, p = 2E-4; downsampling 2 Hz, p = 2E-5; downsampling 5 Hz, p = 8E-9; paired t test n = 13 SLS acquisitions. In this as well in other figures: *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s., non significant. Error bars represent ± SD.