Open-top two-photon light sheet microscopy for three-dimensional tissue examination

Won Yeong Park; Jieun Yun; Jinho Shin; Byung Ho Oh; Gilsuk Yoon; Seung-Mo Hong; Ki Hean Kim

doi:10.1117/12.3016904

26 February 2024 Open-top two-photon light sheet microscopy for three-dimensional tissue examination

Won Yeong Park, Jieun Yun, Jinho Shin, Byung Ho Oh, Gilsuk Yoon, Seung-Mo Hong, Ki Hean Kim

Proceedings Volume 13076, SPIE Advanced Biophotonics Conference (SPIE ABC 2023); 1307603 (2024) https://doi.org/10.1117/12.3016904
Event: SPIE Advanced Biophotonics Conference (SPIE ABC 2023), 2023, Jeju, Korea, Republic of

Abstract

Nondestructive three-dimensional (3D) pathology based on high-throughput 3D microscopy holds promise as a complement to traditional hematoxylin and eosin (H&E) stained slide-based two-dimensional (2D) pathology by providing rapid 3D pathological information. However, conventional techniques provided superficial information only due to shallow imaging depth. Herein, we developed open-top two-photon light sheet microscopy (OT-TPLSM) for intraoperative 3D pathology. A two-photon excitation light sheet, generated by 1D scanning of a Bessel beam illuminated the sample and planar imaging was conducted at 400 frames/s max. An imaging depth of 60-100 μm was achieved with long excitation wavelengths, and the image throughput was up to 1 cm² per 7 min. Cells and extra-cellular matrix were visualized using extrinsic fluorescence and intrinsic second harmonic generation, respectively. OTTPLSM was tested in various human cancer specimens and cancer structures were detected via 3D visualization. OT-TPLSM may have the potential for rapid and precise 3D histopathological examination.

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

Citation Download Citation

Won Yeong Park, Jieun Yun, Jinho Shin, Byung Ho Oh, Gilsuk Yoon, Seung-Mo Hong, and Ki Hean Kim "Open-top two-photon light sheet microscopy for three-dimensional tissue examination", Proc. SPIE 13076, SPIE Advanced Biophotonics Conference (SPIE ABC 2023), 1307603 (26 February 2024); https://doi.org/10.1117/12.3016904

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