Extended Data Fig. 7: Spatial AO enhancement analysis for local aberration correction in-vivo in Thy1-GFP mouse hippocampus.
Aberration measurement was performed for four different conditions referred to: system aberration correction (‘AO system’) and full AO aberration correction inside cortex including Zernike modes up to order 21, (Z21), 35 (Z35) and 60 (Z60), excluding piston, tip and tilt mode. The AO intensity enhancement (AO on/AO off) was then analysed with respect to the distance (lateral and axial) from the point of correction (somata where AO measurement was performed) to estimate the AO isoplanatic patch size (FOV over which the intensity enhancement (AO on/AO off > 1). In total 6 datasets were analysed (total 396 neurons, from 6 image trials in 3 mice). Corresponding aberrations and depth of AO measurements are displayed in Extended Data Fig. 8. a, Lateral (Top, xy) and orthogonal (Bottom, xz) view of maximum intensity projection image of example dataset at 1086-1206μm depth across 360×360μm lateral FOV acquired without (AO off) and full (AO on Z35) aberration correction. Scale bar 20μm. b, Instance segmentation of neuron somata in (a). c, Relative position of centroids of all segmented somata (396 neurons) pooled from 6 datasets with respect to point of correction (x=0, y=0, z=0). d, Spatial projection of somata (all 6 datasets) with respect to lateral (Rxy) and axial (Rz) distance from point of correction (Rxy =0, Rz =0). Colour coding represents intensity enhancement (AO on/ AO off) where red colour is >1, and green <1. e, Lateral and axial data from (c) grouped into distance increments of 50 µm. For lateral analysis only somata with Rz < 50 µm and accordingly for the axial analysis only somata with Rxy< 50 µm where analyzed. Boxplot shows median value as red line. Boundaries of box represent upper 75th and lower 25th percentile, whiskers correspond to +/–2.7σ and 99.3 percent coverage. The region where intensity enhancement (AO on/AO off) is <1 is indicated by red box. Analysis shows that the isoplanatic patch (AO on/AO off > 1) is larger in the axial compared to the lateral dimension. Summary plot for Z35 is included in Fig. 2h.
