Single-objective light-sheet microscopy enables multiscale imaging spanning from subcellular structures to whole organs

Yao Zhou; Xinyi Guo; Shiqi Mao; Jianchao Liu; Peng Fei

doi:10.1117/12.3047959

13 December 2024 Single-objective light-sheet microscopy enables multiscale imaging spanning from subcellular structures to whole organs

Yao Zhou, Xinyi Guo, Shiqi Mao, Jianchao Liu, Peng Fei

Author Affiliations +

Proceedings Volume 13503, AOPC 2024: Biomedical Optics; 135030B (2024) https://doi.org/10.1117/12.3047959
Event: Applied Optics and Photonics China 2024 (AOPC2024), 2024, Beijing, China

Abstract

Light-sheet fluorescence microscopy (LSFM) is an advanced three-dimensional microscopic imaging technique, possessing unique imaging advantages for samples of various scales. One the one hand, for mesoscale samples, its tomographic sheet-like illumination allows the acquisition of information from a whole plane at a time, greatly increasing the imaging throughput (the amount of information obtained per unit time). On the other hand, for nanoscale samples, its three-dimensional resolution and photobleaching performance are both superior to those of traditional widefield illumination optical microscopes. However, traditional LSFM's dual-objective setup limits mounting options and speed. Single-objective LSFM (OPM) addresses this, but still has drawbacks like limited field of view and need for additional objectives. Addressing these limitations is crucial for LSFM's full potential in biomedical research. Herein, we propose double-beam interference Microscopy, a novel single-objective LSFM design which could realize horizontal plane illumination using two oblique laser sheets, compatible with various sample mounts. It eliminates remote correction, enabling imaging from organelles to mouse brains by switching objectives. Its sharp optical sectioning and full-aperture detection offer superior spatiotemporal resolution. We showcase its versatility in neuroanatomy and immunology imaging, enabling coarse-to-fine imaging, automatic coordinate mapping, and anatomical annotation correlation.

(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

Citation Download Citation

Yao Zhou, Xinyi Guo, Shiqi Mao, Jianchao Liu, and Peng Fei "Single-objective light-sheet microscopy enables multiscale imaging spanning from subcellular structures to whole organs", Proc. SPIE 13503, AOPC 2024: Biomedical Optics, 135030B (13 December 2024); https://doi.org/10.1117/12.3047959

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KEYWORDS

3D image processing

Imaging systems

Biological imaging

Microscopy

Fluorescence imaging

Fluorescence

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