格廷根大学Jan Huisken研究组报道了用于清除组织的快速高分辨率成像的各向同性、像差校正光片显微镜。该研究于2025年11月13日发表于国际一流学术期刊《自然—生物技术》杂志上。

研究小组介绍了一种紧凑、高速的光片荧光显微镜，在清除样品上实现850 nm各向同性分辨率，最高可达1 cm³，折射率范围为1.33至1.56。使用现成的光学器件，该研究组将一个空中物镜和一个半月板透镜与一个轴向扫光片相结合，以实现衍射极限分辨率和像差校正。通过对远距调焦单元内凹面镜光片的场曲率进行校正，使有效视场增加了两倍。通过闭环反馈调整光片的运动，可以将成像速度提高10倍，在保持分辨率和视野的同时达到每秒100帧。该研究团队对系统性能进行了跨尺度的基准测试，从亚细胞结构到厘米尺度，主题为各种清除方法。

据介绍，光片显微镜是理想的成像大和清除组织，但实现高各向同性分辨率为厘米大小的样品是受限制的缓慢和经常畸变，轴向扫描光片。

附：英文原文

Title: Isotropic, aberration-corrected light sheet microscopy for rapid high-resolution imaging of cleared tissue

Author: Aakhte, Mostafa, Mller, Gesine F., Roos, Lennart, Li, Joe, Gpel, Torben, Weiss, Kurt R., Diniz, Aleyna M., Wenzel, Jan, Schwaninger, Markus, Moser, Tobias, Huisken, Jan

Issue&Volume: 2025-11-13

Abstract: Light-sheet microscopy is ideal for imaging large and cleared tissues, but achieving a high isotropic resolution for a centimeter-sized sample is limited by slow and often aberrated, axially scanned light sheets. Here, we introduce a compact, high-speed light-sheet fluorescence microscope achieving 850 nm isotropic resolution across cleared samples up to 1 cm3 and refractive indices ranging from 1.33 to 1.56. Using off-the-shelf optics, we combine an air objective and a meniscus lens with an axially swept light sheet to achieve diffraction-limited resolution and aberration correction. The effective field of view is increased by twofold by correcting the field curvature of the light sheet using a concave mirror in the remote focusing unit. Adapting the light sheet’s motion with a closed-loop feedback enhances the imaging speed by tenfold, reaching 100 frames per second while maintaining resolution and field of view. We benchmark the system performance across scales, from subcellular structure up to centimeter scale, using various clearing methods.

DOI: 10.1038/s41587-025-02882-8

Source: https://www.nature.com/articles/s41587-025-02882-8