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Open-top Bessel beam two-photon light sheet microscopy for three-dimensional pathology

 Won Yeong Park  ;  Jieun Yun  ;  Jinho Shin  ;  Byung Ho Oh  ;  Gilsuk Yoon  ;  Seung-Mo Hong  ;  Ki Hean Kim 
 ELIFE, Vol.12 : RP92614, 2024-03 
Journal Title
Issue Date
Eosine Yellowish-(YS) ; Fluorescent Dyes ; Humans ; Imaging, Three-Dimensional / methods ; Male ; Microscopy* / methods ; Neoplasms* ; Skin
3D imaging ; 3D pathology ; bessel beam illumination ; cancer biology ; human ; medicine ; nondestructive pathology ; two-photon light sheet microscopy ; unsupervised deep learning
Nondestructive pathology based on three-dimensional (3D) optical microscopy holds promise as a complement to traditional destructive hematoxylin and eosin (H&E) stained slide-based pathology by providing cellular information in high throughput manner. However, conventional techniques provided superficial information only due to shallow imaging depths. Herein, we developed open-top two-photon light sheet microscopy (OT-TP-LSM) for intraoperative 3D pathology. An extended depth of field two-photon excitation light sheet was generated by scanning a nondiffractive Bessel beam, and selective planar imaging was conducted with cameras at 400 frames/s max during the lateral translation of tissue specimens. Intrinsic second harmonic generation was collected for additional extracellular matrix (ECM) visualization. OT-TP-LSM was tested in various human cancer specimens including skin, pancreas, and prostate. High imaging depths were achieved owing to long excitation wavelengths and long wavelength fluorophores. 3D visualization of both cells and ECM enhanced the ability of cancer detection. Furthermore, an unsupervised deep learning network was employed for the style transfer of OT-TP-LSM images to virtual H&E images. The virtual H&E images exhibited comparable histological characteristics to real ones. OT-TP-LSM may have the potential for histopathological examination in surgical and biopsy applications by rapidly providing 3D information. © 2023, Park et al.
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1. College of Medicine (의과대학) > Dept. of Dermatology (피부과학교실) > 1. Journal Papers
Yonsei Authors
Oh, Byung Ho(오병호) ORCID logo https://orcid.org/0000-0001-9575-5665
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