Cryo-focused ion beam milling for cryo-electron tomography: Shaping the future of in situ structural biology

IF 6.1 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Current opinion in structural biology Pub Date : 2025-08-22 DOI:10.1016/j.sbi.2025.103138

Sven Klumpe , Jürgen M. Plitzko

引用次数: 0

Abstract

Cryo-focused ion beam instruments to produce cellular thin sections for subsequent imaging by cryo-electron tomography have become an integral part of the methodologies for in situ structural biology, enabling high-resolution imaging of biological structures in their native environment. The application of these instruments has opened windows into cells that allowed unprecedented insights into the ultrastructure of cells and more recently, small multicellular organisms and tissues. While great strides have been made in the characterization of cryo-FIB milling and the streamlining of workflows with these tools, many limitations and technical challenges remain to be resolved. Here, we discuss the technical and technological challenges ahead to continue the steep rise of biological discoveries by in-cell cryo-electron tomography to enable cellular structural biology in the multicellular context.

查看原文本刊更多论文

低温聚焦离子束铣削用于低温电子断层扫描：塑造原位结构生物学的未来

低温聚焦离子束仪器为随后的低温电子断层成像生产细胞薄切片，已经成为原位结构生物学方法的一个组成部分，使生物结构在其原生环境中的高分辨率成像成为可能。这些仪器的应用为研究细胞打开了一扇窗，使我们能够前所未有地深入了解细胞的超微结构，以及最近的小型多细胞生物和组织。虽然在低温fib铣削表征和使用这些工具简化工作流程方面取得了很大进展，但仍有许多限制和技术挑战有待解决。在这里，我们讨论了未来的技术和技术挑战，以继续通过细胞内低温电子断层扫描来实现多细胞环境下的细胞结构生物学的生物学发现。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Current opinion in structural biology 生物-生化与分子生物学

CiteScore

12.20

自引率

2.90%

发文量

179

审稿时长

6-12 weeks

期刊介绍： Current Opinion in Structural Biology (COSB) aims to stimulate scientifically grounded, interdisciplinary, multi-scale debate and exchange of ideas. It contains polished, concise and timely reviews and opinions, with particular emphasis on those articles published in the past two years. In addition to describing recent trends, the authors are encouraged to give their subjective opinion of the topics discussed. In COSB, we help the reader by providing in a systematic manner: 1. The views of experts on current advances in their field in a clear and readable form. 2. Evaluations of the most interesting papers, annotated by experts, from the great wealth of original publications. [...] The subject of Structural Biology is divided into twelve themed sections, each of which is reviewed once a year. Each issue contains two sections, and the amount of space devoted to each section is related to its importance. -Folding and Binding- Nucleic acids and their protein complexes- Macromolecular Machines- Theory and Simulation- Sequences and Topology- New constructs and expression of proteins- Membranes- Engineering and Design- Carbohydrate-protein interactions and glycosylation- Biophysical and molecular biological methods- Multi-protein assemblies in signalling- Catalysis and Regulation