Tilt-invariant scanned oblique plane illumination microscopy for large-scale volumetric imaging

This Letter presents the first demonstration of multi-tile stitching for large scale 3D imaging in single objective light-sheet microscopy. We show undistorted 3D imaging spanning complete zebrafish larvae and over 1  mm3 volumes for thick mouse brain sections. We use remote galvo scanning for light-sheet creation and develop a processing pipeline for 3D tiling across different axes. With the improved one photon (1p) tilt-invariant scanned oblique plane illumination (SOPi, /sōpī/) microscope presented here, we demonstrate cellular resolution imaging at depths exceeding 330 μm in optically scattering mouse brain samples and dendritic imaging in more superficial layers.