PDF(Pubmed)
Abstract：
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.
摘要：
基于三维（3D）光学显微镜的无损病理学有望通过以高通量方式提供细胞信息来补充传统的破坏性苏木精和伊红（H＆E）染色的基于载玻片的病理学。然而,传统技术仅由于浅层成像深度而提供浅层信息。在这里，我们开发了用于术中3D病理的开放式双光子光片显微镜（OT-TP-LSM）。通过扫描非衍射贝塞尔光束，产生了扩展的景深双光子激发光片，在组织标本的横向平移过程中，用摄像机以最大400帧/s的速度进行选择性平面成像。收集固有二次谐波产生用于额外的细胞外基质(ECM)可视化。OT-TP-LSM在包括皮肤在内的各种人类癌症标本中进行了测试，胰腺,还有前列腺.由于长激发波长和长波长荧光团，可以实现高成像深度。细胞和ECM的3D可视化增强了癌症检测的能力。此外，采用无监督的深度学习网络将OT-TP-LSM图像样式转换为虚拟H&E图像。虚拟H＆E图像表现出与真实图像相当的组织学特征。OT-TP-LSM可能通过快速提供3D信息而在手术和活检应用中具有组织病理学检查的潜力。
