Integrated one- and two-photon scanned oblique plane illumination (SOPi) microscopy for rapid volumetric imaging

TL;DR

This paper introduces an integrated SOPi microscopy system combining one- and two-photon light-sheet imaging for rapid, high-resolution volumetric imaging of brain tissue and live organisms, enhancing neuroscience research capabilities.

Contribution

The novel SOPi system uniquely integrates one- and two-photon microscopy using a single objective, enabling versatile, rapid volumetric imaging with simplified reconstruction in scattering media.

Findings

Deep imaging in scattering mouse brain sections

Rapid imaging of zebrafish larvae at up to 10 volumes/sec

Compatible with various neuronal activity indicators

Abstract

Versatile, sterically accessible imaging systems capable of in vivo rapid volumetric functional and structural imaging deep in the brain continue to be a limiting factor in neuroscience research. Towards overcoming this obstacle, we present integrated one- and two-photon scanned oblique plane illumination (SOPi) microscopy which uses a single front-facing microscope objective to provide light-sheet scanning based rapid volumetric imaging capability at subcellular resolution. Our planar scan-mirror based optimized light-sheet architecture allows for non-distorted scanning of volume samples, simplifying accurate reconstruction of the imaged volume. Integration of both one-photon (1P) and two-photon (2P) light-sheet microscopy in the same system allows for easy selection between rapid volumetric imaging and higher resolution imaging in scattering media. Using SOPi, we demonstrate deep, large volume imaging capability inside scattering mouse brain sections and rapid imaging speeds up to 10 volumes per second in zebrafish larvae expressing genetically encoded fluorescent proteins GFP or GCaMP6s. SOPi flexibility and steric access makes it adaptable for numerous imaging applications and broadly compatible with orthogonal techniques for actuating or interrogating neuronal structure and activity.