Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang 310024, China; Institute of Basic Medical Sciences, Westlake Institute for Advanced Study, Hangzhou, Zhejiang 310024, China; Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang 310024, China.
Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang 310024, China; Institute of Basic Medical Sciences, Westlake Institute for Advanced Study, Hangzhou, Zhejiang 310024, China.
Cell Rep. 2020 Nov 3;33(5):108349. doi: 10.1016/j.celrep.2020.108349.
We present a tiling light sheet microscope compatible with all tissue clearing methods for rapid multicolor 3D imaging of cleared tissues with micron-scale (4 × 4 × 10 μm) to submicron-scale (0.3 × 0.3 × 1 μm) spatial resolution. The resolving ability is improved to sub-100 nm (70 × 70 × 200 nm) via tissue expansion. The microscope uses tiling light sheets to achieve higher spatial resolution and better optical sectioning ability than conventional light sheet microscopes. The illumination light is phase modulated to adjust the position and intensity profile of the light sheet based on the desired spatial resolution and imaging speed and to keep the microscope aligned. The ability of the microscope to align via phase modulation alone also ensures its accuracy for multicolor 3D imaging and makes the microscope reliable and easy to operate. Here we describe the working principle and design of the microscope. We demonstrate its utility by imaging various cleared tissues.
我们提出了一种适用于所有组织透明化方法的平铺光片显微镜，可用于快速对透明组织进行多色 3D 成像，空间分辨率达到亚微米级（4μm×4μm×10μm）至亚 100nm 级（70nm×70nm×200nm）。显微镜通过组织扩展，将分辨率提高到亚 100nm 级。该显微镜采用平铺光片，与传统的光片显微镜相比，具有更高的空间分辨率和更好的光学切片能力。照明光采用相位调制，根据所需的空间分辨率和成像速度来调整光片的位置和强度分布，并保持显微镜的对齐。显微镜仅通过相位调制进行对齐的能力也确保了其进行多色 3D 成像的准确性，使其具有可靠性和易于操作的特点。本文描述了该显微镜的工作原理和设计。我们通过对各种透明组织进行成像来证明其用途。
