小柯机器人

中国科学院Wei Ji，Tao Xu和北京大学Qiang Guo共同合作，近期取得重要工作进展。他们研究开发了集成多模显微镜用于精确和高效的目标导向冷冻薄片制备。相关研究成果2023年1月16日在线发表于《自然—方法学》杂志上。

据介绍，冷冻电子断层扫描（Cryo-ET）是一种革命性的技术，用于在细胞环境中解析亚细胞细胞器和大分子复合物的结构。然而，Cryo-ET的应用受到样品制备步骤的阻碍。在样品上的任意位置执行低温聚焦离子束铣削是低效的，并且当将细胞从几微米减薄到小于300纳米厚时，不能保证保留感兴趣的目标位置。

研究人员开发了一种低温相关光、离子和电子显微镜（cryo-CLIEM）技术，该技术能够在三维共焦成像的指导下制备冷冻薄片。

此外，研究人员演示了在完成的冷冻薄片内预选和保存纳米级目标区域的工作流程。通过成功制备包含单个中心粒或亚细胞细胞器之间的接触位点的冷冻薄片，研究人员表明这种方法是普遍适用的，并将有助于开发cryo-ET的更多应用。

附：英文原文

Title: Integrated multimodality microscope for accurate and efficient target-guided cryo-lamellae preparation

Author: Li, Weixing, Lu, Jing, Xiao, Ke, Zhou, Maoge, Li, Yuanyuan, Zhang, Xiang, Li, Zhixun, Gu, Lusheng, Xu, Xiaojun, Guo, Qiang, Xu, Tao, Ji, Wei

Issue&Volume: 2023-01-16

Abstract: Cryo-electron tomography (cryo-ET) is a revolutionary technique for resolving the structure of subcellular organelles and macromolecular complexes in their cellular context. However, the application of the cryo-ET is hampered by the sample preparation step. Performing cryo-focused ion beam milling at an arbitrary position on the sample is inefficient, and the target of interest is not guaranteed to be preserved when thinning the cell from several micrometers to less than 300nm thick. Here, we report a cryogenic correlated light, ion and electron microscopy (cryo-CLIEM) technique that is capable of preparing cryo-lamellae under the guidance of three-dimensional confocal imaging. Moreover, we demonstrate a workflow to preselect and preserve nanoscale target regions inside the finished cryo-lamellae. By successfully preparing cryo-lamellae that contain a single centriole or contact sites between subcellular organelles, we show that this approach is generally applicable, and shall help in innovating more applications of cryo-ET.

DOI: 10.1038/s41592-022-01749-z

Source: https://www.nature.com/articles/s41592-022-01749-z

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