Villegas-Hernández Luis E, Dubey Vishesh, Nystad Mona, Tinguely Jean-Claude, Coucheron David A, Dullo Firehun T, Priyadarshi Anish, Acuña Sebastian, Ahmad Azeem, Mateos José M, Barmettler Gery, Ziegler Urs, Birgisdottir Åsa Birna, Hovd Aud-Malin Karlsson, Fenton Kristin Andreassen, Acharya Ganesh, Agarwal Krishna, Ahluwalia Balpreet Singh
Department of Physics and Technology, UiT The Arctic University of Norway, Klokkargårdsbakken N-9019, Tromsø, Norway.
Department of Clinical Medicine, Women's Health and Perinatology Research Group, UiT The Arctic University of Norway, Tromsø, Norway.
Light Sci Appl. 2022 Feb 24;11(1):43. doi: 10.1038/s41377-022-00731-w.
Histology involves the observation of structural features in tissues using a microscope. While diffraction-limited optical microscopes are commonly used in histological investigations, their resolving capabilities are insufficient to visualize details at subcellular level. Although a novel set of super-resolution optical microscopy techniques can fulfill the resolution demands in such cases, the system complexity, high operating cost, lack of multi-modality, and low-throughput imaging of these methods limit their wide adoption for histological analysis. In this study, we introduce the photonic chip as a feasible high-throughput microscopy platform for super-resolution imaging of histological samples. Using cryopreserved ultrathin tissue sections of human placenta, mouse kidney, pig heart, and zebrafish eye retina prepared by the Tokuyasu method, we demonstrate diverse imaging capabilities of the photonic chip including total internal reflection fluorescence microscopy, intensity fluctuation-based optical nanoscopy, single-molecule localization microscopy, and correlative light-electron microscopy. Our results validate the photonic chip as a feasible imaging platform for tissue sections and pave the way for the adoption of super-resolution high-throughput multimodal analysis of cryopreserved tissue samples both in research and clinical settings.
组织学涉及使用显微镜观察组织中的结构特征。虽然衍射极限光学显微镜在组织学研究中常用，但其分辨能力不足以在亚细胞水平可视化细节。尽管一组新颖的超分辨率光学显微镜技术能够满足此类情况下的分辨率要求，但这些方法的系统复杂性、高运行成本、缺乏多模态以及低通量成像限制了它们在组织学分析中的广泛应用。在本研究中，我们引入光子芯片作为用于组织学样本超分辨率成像的可行高通量显微镜平台。使用通过德永方法制备的人胎盘、小鼠肾脏、猪心脏和斑马鱼眼视网膜的冷冻超薄组织切片，我们展示了光子芯片的多种成像能力，包括全内反射荧光显微镜、基于强度波动的光学纳米显微镜、单分子定位显微镜和相关光电子显微镜。我们的结果验证了光子芯片作为组织切片可行成像平台的有效性，并为在研究和临床环境中采用冷冻保存组织样本的超分辨率高通量多模态分析铺平了道路。
