Kim Laura Y, Rice William J, Eng Edward T, Kopylov Mykhailo, Cheng Anchi, Raczkowski Ashleigh M, Jordan Kelsey D, Bobe Daija, Potter Clinton S, Carragher Bridget
Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, United States.
Front Mol Biosci. 2018 Jun 4;5:50. doi: 10.3389/fmolb.2018.00050. eCollection 2018.
Cryo electron microscopy facilities running multiple instruments and serving users with varying skill levels need a robust and reliable method for benchmarking both the hardware and software components of their single particle analysis workflow. The workflow is complex, with many bottlenecks existing at the specimen preparation, data collection and image analysis steps; the samples and grid preparation can be of unpredictable quality, there are many different protocols for microscope and camera settings, and there is a myriad of software programs for analysis that can depend on dozens of settings chosen by the user. For this reason, we believe it is important to benchmark the entire workflow, using a standard sample and standard operating procedures, on a regular basis. This provides confidence that all aspects of the pipeline are capable of producing maps to high resolution. Here we describe benchmarking procedures using a test sample, rabbit muscle aldolase.
运行多台仪器并为不同技能水平的用户提供服务的冷冻电子显微镜设施，需要一种强大且可靠的方法来对其单颗粒分析工作流程的硬件和软件组件进行基准测试。该工作流程很复杂，在样品制备、数据采集和图像分析步骤存在许多瓶颈；样品和网格制备的质量可能不可预测，显微镜和相机设置有许多不同的协议，并且有大量用于分析的软件程序，这些程序可能取决于用户选择的数十种设置。因此，我们认为定期使用标准样品和标准操作程序对整个工作流程进行基准测试很重要。这让人相信该流程的所有方面都能够生成高分辨率的图谱。在这里，我们描述了使用测试样品兔肌肉醛缩酶的基准测试程序。
