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

Abstract

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) microscope presented here, we demonstrate cellular resolution imaging at depths exceeding 330 um in optically scattering mouse brain samples, and dendritic imaging in more superficial layers.