Fislage Marcus, Shkumatov Alexander V, Stroobants Annelore, Efremov Rouslan G
Center for Structural Biology, Vlaams Instituut voor Biotechnologie, Pleinlaan 2, Brussels 1050, Belgium.
Structural Biology Brussels, Department of Bioengineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, Brussels 1050, Belgium.
IUCrJ. 2020 Jun 11;7(Pt 4):707-718. doi: 10.1107/S2052252520006065. eCollection 2020 Jul 1.
Single-particle cryo-EM has become an indispensable structural biology method. It requires regular access to high-resolution electron cryogenic microscopes. To fully utilize the capacity of the expensive high-resolution instruments, the time used for data acquisition and the rate of data collection have to be maximized. This in turn requires high stability and high uptime of the instrument. One of the first 300 kV JEOL CRYO ARM 300 microscopes has been installed at the cryo-EM facility BECM at VIB-VUB, Brussels, where the microscope is used for continuous data collection on multiple projects. Here, the suitability and performance of the microscope is assessed for high-throughput single-particle data collection. In particular, the properties of the illumination system, the stage stability and ice contamination rates are reported. The microscope was benchmarked using mouse heavy-chain apoferritin which was reconstructed to a resolution of 1.9 Å. Finally, uptime and throughput statistics of the instrument accumulated during the first six months of the facility operation in user access mode are reported.
单颗粒冷冻电镜已成为一种不可或缺的结构生物学方法。它需要定期使用高分辨率电子低温显微镜。为了充分利用昂贵的高分辨率仪器的性能,必须最大限度地提高数据采集时间和数据收集速率。这反过来又要求仪器具有高稳定性和高正常运行时间。首台300 kV的日本电子CRYO ARM 300显微镜已安装在布鲁塞尔VIB - VUB的冷冻电镜设施BECM中,该显微镜用于多个项目的连续数据采集。在此,评估了该显微镜用于高通量单颗粒数据采集的适用性和性能。特别报告了照明系统的特性、样品台稳定性和冰污染率。使用小鼠重链脱铁铁蛋白对该显微镜进行了基准测试,其重构分辨率达到了1.9 Å。最后,报告了该仪器在设施用户访问模式运行的前六个月积累的正常运行时间和通量统计数据。