Single-objective selective-volume illumination microscopy enables high-contrast light-field imaging

TL;DR

This paper introduces a single-objective selective-volume illumination microscopy technique using oblique or two-photon illumination, enhancing light-field imaging by improving contrast and resolution without requiring a second objective.

Contribution

It advances light-field microscopy by enabling selective-volume excitation in a single-objective setup, simplifying specimen mounting and reducing background noise.

Findings

Improved image contrast and resolution in light-field microscopy.

Successful imaging of live zebrafish with high contrast.

Compatible with standard microscope sample mounting.

Abstract

The performance of light-field microscopy is improved by selectively illuminating the relevant subvolume of the specimen with a second objective lens [1-3]. Here we advance this approach to a single-objective geometry, using an oblique one-photon illumination path or two-photon illumination to accomplish selective-volume excitation. The elimination of the second orthogonally oriented objective to selectively excite the volume of interest simplifies specimen mounting; yet, this single-objective approach still reduces out-of-volume background, resulting in improvements in image contrast, effective resolution, and volume reconstruction quality. We validate our new approach through imaging live developing zebrafish, demonstrating the technology's ability to capture imaging data from large volumes synchronously with high contrast, while remaining compatible with standard microscope sample mounting.