小柯机器人

日本东京大学Radostin Danev研究团队报道并行低温电子断层扫描在原位薄层上的应用。2022年12月1日，国际知名学术期刊《自然—方法学》在线发表了这一成果。

研究人员利用一个几何样品模型和光学图像移位来平行记录了几十个倾斜系列，从而节省时间，并获得传统上用于跟踪试样运动的样品区域。并行低温电子断层扫描（PACE-tomo）方法实现了每个倾斜系列快于5分钟的吞吐量，并允许收集以前无法到达的样品区域，从而最大限度地提高每个薄层的数据量。在最先进的和通用的显微镜上用核糖体在体外和原位进行的性能测试证明了PACE-tomo的高通量和质量。

据了解，近年来，低温聚焦离子束研磨样品的原位低温电子断层成像技术已经成为一种强大的技术，被用于对大分子复合物在其原始细胞环境中的结构研究。然而，以高通量的方式记录断层扫描系列的可能性是有限的，部分原因是片状的样品。

附：英文原文

Title: Parallel cryo electron tomography on in situ lamellae

Author: Eisenstein, Fabian, Yanagisawa, Haruaki, Kashihara, Hiroka, Kikkawa, Masahide, Tsukita, Sachiko, Danev, Radostin

Issue&Volume: 2022-12-01

Abstract: In situ cryo electron tomography of cryo focused ion beam milled samples has emerged in recent years as a powerful technique for structural studies of macromolecular complexes in their native cellular environment. However, the possibilities for recording tomographic tilt series in a high-throughput manner are limited, in part by the lamella-shaped samples. Here we utilize a geometrical sample model and optical image shift to record tens of tilt series in parallel, thereby saving time and gaining access to sample areas conventionally used for tracking specimen movement. The parallel cryo electron tomography (PACE-tomo) method achieves a throughput faster than 5min per tilt series and allows for the collection of sample areas that were previously unreachable, thus maximizing the amount of data from each lamella. Performance testing with ribosomes in vitro and in situ on state-of-the-art and general-purpose microscopes demonstrated the high throughput and quality of PACE-tomo.

DOI: 10.1038/s41592-022-01690-1

Source: https://www.nature.com/articles/s41592-022-01690-1

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