小柯机器人

美国德克萨斯大学西南医学中心Reto P. Fiolka小组近日取得一项新成果。经过不懈努力，他们通过斜平面结构照明实现了光片显微镜分辨率的加倍。该项研究成果发表在2022年10月24日出版的《自然-方法学》上。

研究人员展示了多向结构化照明可以在倾斜平面显微镜中实现，这是一种光片荧光显微镜（LSFM）技术，它使用单个物镜以直接的方式进行激发和检测。研究展示了低于150 nm的各向同性横向分辨率，与传统SIM系统相比，光毒性更低并且体积采集速度超过1 Hz。

据介绍，结构化照明显微镜（SIM）的空间分辨率是荧光显微镜的两倍，而且其无需高激光功率或专用荧光团。然而，失焦荧光的激发会加速光漂白和光毒性。相比之下， LSFM在很大程度上避免了激发失焦荧光，从而实现了低光漂白和固有光学切片体积成像。将SIM与LSFM相结合，可以双倍分辨率实现柔和三维（3D）成像。然而，SIM实现各向同性分辨率加倍所需的照明模式在多个方向具有挑战，很难以光片格式实现。

附：英文原文

Title: Resolution doubling in light-sheet microscopy via oblique plane structured illumination

Author: Chen, Bingying, Chang, Bo-Jui, Roudot, Philippe, Zhou, Felix, Sapoznik, Etai, Marlar-Pavey, Madeleine, Hayes, James B., Brown, Peter T., Zeng, Chih-Wei, Lambert, Talley, Friedman, Jonathan R., Zhang, Chun-Li, Burnette, Dylan T., Shepherd, Douglas P., Dean, Kevin M., Fiolka, Reto P.

Issue&Volume: 2022-10-24

Abstract: Structured illumination microscopy (SIM) doubles the spatial resolution of a fluorescence microscope without requiring high laser powers or specialized fluorophores. However, the excitation of out-of-focus fluorescence can accelerate photobleaching and phototoxicity. In contrast, light-sheet fluorescence microscopy (LSFM) largely avoids exciting out-of-focus fluorescence, thereby enabling volumetric imaging with low photobleaching and intrinsic optical sectioning. Combining SIM with LSFM would enable gentle three-dimensional (3D) imaging at doubled resolution. However, multiple orientations of the illumination pattern, which are needed for isotropic resolution doubling in SIM, are challenging to implement in a light-sheet format. Here we show that multidirectional structured illumination can be implemented in oblique plane microscopy, an LSFM technique that uses a single objective for excitation and detection, in a straightforward manner. We demonstrate isotropic lateral resolution below 150nm, combined with lower phototoxicity compared to traditional SIM systems and volumetric acquisition speed exceeding 1Hz.

DOI: 10.1038/s41592-022-01635-8

Source: https://www.nature.com/articles/s41592-022-01635-8

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex