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

TL;DR

This paper introduces a tilt-invariant scanned oblique plane illumination microscopy technique enabling large-scale, high-resolution 3D imaging of biological samples like zebrafish larvae and mouse brains, using multi-tile stitching and remote galvo scanning.

Contribution

It presents the first implementation of multi-tile stitching in single objective light-sheet microscopy with tilt-invariant illumination for large-volume imaging.

Findings

Achieved undistorted 3D imaging of entire zebrafish larvae.

Imaged over 1 mm³ volumes of mouse brain tissue.

Demonstrated cellular resolution at depths exceeding 330 μm in scattering tissue.

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.