Materials and Structural Analysis, Thermo Fisher Scientific, Achtseweg Noord 5, 5651 GG Eindhoven, The Netherlands.
Acta Crystallogr D Struct Biol. 2020 Apr 1;76(Pt 4):313-325. doi: 10.1107/S2059798320002223. Epub 2020 Apr 6.
Cryo-electron microscopy (cryo-EM) has rapidly expanded with the introduction of direct electron detectors, improved image-processing software and automated image acquisition. Its recent adoption by industry, particularly in structure-based drug design, creates new requirements in terms of reliability, reproducibility and throughput. In 2016, Thermo Fisher Scientific (then FEI) partnered with the Medical Research Council Laboratory of Molecular Biology, the University of Cambridge Nanoscience Centre and five pharmaceutical companies [Astex Pharmaceuticals, AstraZeneca, GSK, Sosei Heptares and Union Chimique Belge (UCB)] to form the Cambridge Pharmaceutical Cryo-EM Consortium to share the risks of exploring cryo-EM for early-stage drug discovery. The Consortium expanded with a second Themo Scientific Krios Cryo-EM at the University of Cambridge Department of Materials Science and Metallurgy. Several Consortium members have set up in-house facilities, and a full service cryo-EM facility with Krios and Glacios has been created with the Electron Bio-Imaging Centre for Industry (eBIC for Industry) at Diamond Light Source (DLS), UK. This paper will cover the lessons learned during the setting up of these facilities, including two Consortium Krios microscopes and preparation laboratories, several Glacios microscopes at Consortium member sites, and a Krios and Glacios at eBIC for Industry, regarding site evaluation and selection for high-resolution cryo-EM microscopes, the installation process, scheduling, the operation and maintenance of the microscopes and preparation laboratories, and image processing.
冷冻电子显微镜（cryo-EM）随着直接电子探测器的引入、图像处理软件的改进和自动化图像采集技术的发展而迅速发展。其最近在工业界的应用，特别是在基于结构的药物设计中，对可靠性、可重复性和通量提出了新的要求。2016 年，赛默飞世尔科技（当时的 FEI）与英国剑桥分子生物学实验室、剑桥纳米科学中心和五家制药公司[Astex 制药、阿斯利康、葛兰素史克、Sosei Heptares 和 Union Chimique Belge (UCB)]合作成立了剑桥制药冷冻电子显微镜联盟，共同承担探索冷冻电子显微镜在早期药物发现中的风险。该联盟在剑桥大学材料科学与冶金系又增加了一台赛默飞世尔科技 Krios 冷冻电子显微镜。一些联盟成员已经建立了内部设施，并与英国钻石光源公司的电子生物成像中心（eBIC for Industry）合作，建立了一个拥有 Krios 和 Glacios 的全方位服务冷冻电子显微镜设施。本文将介绍在这些设施建设过程中获得的经验教训，包括两台联盟 Krios 显微镜和准备实验室、联盟成员现场的几台 Glacios 显微镜，以及 eBIC for Industry 的 Krios 和 Glacios，内容涉及高分辨率冷冻电子显微镜的站点评估和选择、安装过程、调度、显微镜和准备实验室的操作和维护，以及图像处理。
