300 kV Cryo-Transmission Electron Microscope for tomography

在生命科学中，从分子到细胞的生物对象的可视化和结构确定对于在空间和时间上分析生理和病理生理过程至关重要。超分子组装体或功能模块对于生物功能和组织特别感兴趣，因为它们整合分子反应和相互作用以产生生物复杂性。超分子组装体和大分子机器的结构分析的一个关键方法是三维冷冻电子显微镜（3D cryo-EM）。最近的革命性技术突破使得（i）通过单粒子以近原子分辨率确定纯化的大分子组装体的结构和（ii）用低温电子断层扫描（cryo-ET）以纳米至亚纳米分辨率可视化细胞的天然环境中的超分子组装体成为可能。为了在柏林建立细胞冷冻电镜，需要一台具有相位板、能量过滤器和直接电子探测器的高端300 kV冷冻电镜仪器。该设置也适用于具有挑战性的较小复合物（< 300 kDa）的单颗粒冷冻EM。该仪器应集成到Charité EM核心设施中，并与该组合应用的其他仪器一起集成到柏林范围内的先进冷冻EM网络中。所要求的仪器应确保柏林的这一开创性技术，促进新的综合结构生物学方法，并作为相关和多尺度成像的中心。

In life sciences, visualization and structure determination of biological objects from molecules to cells is of crucial importance for the analysis of physiologic and pathophysiologic processes in space and time. Supramolecular assemblies or functional modules are of particular interest for biological function and organization as they integrate molecular reactions and interactions to generate biological complexity. A key method for the structural analysis of supramolecular assemblies and macromolecular machines is threedimensional cryo-electron microscopy (3D cryo-EM). Recent revolutionary technological breakthroughs make it possible (i) to determine the structure of purified macromolecular assemblies at near -atomic resolution by single-particle and (ii) to visualize supramolecular assemblies in the native environment of the cell at nanometer to sub-nanometer resolution with cryo-electron tomography (cryo-ET). To establish cellular cryo-ET in Berlin, a high-end 300 kV cryo-EM instrument with phase plate, energy filter and direct electron detector is required. This setup is also suited for single-particle cryo-EM of challenging, smaller complexes (< 300 kDa). The instrument shall be integrated into the Charité EM core facility and, together with the other instruments of this combined application, into a Berlin-wide network for advanced cryo-EM. The requested instrumentation shall secure this seminal technology for Berlin, facilitate novel integrated structural biology approaches, and serve as a hub for correlative and multiscale imaging.