PDF(Pubmed)
Abstract：
In addressing the challenges faced by laboratories and universities with limited (or no) cryo-electron microscopy (cryo-EM) infrastructure, the ESRF, in collaboration with the Grenoble Institute for Structural Biology (IBS), has implemented the cryo-EM Solution-to-Structure (SOS) pipeline. This inclusive process, spanning grid preparation to high-resolution data collection, covers single-particle analysis and cryo-electron tomography (cryo-ET). Accessible through a rolling access route, proposals undergo scientific merit and technical feasibility evaluations. Stringent feasibility criteria demand robust evidence of sample homogeneity. Two distinct entry points are offered: users can either submit purified protein samples for comprehensive processing or initiate the pipeline with already vitrified cryo-EM grids. The SOS pipeline integrates negative stain imaging (exclusive to protein samples) as a first quality step, followed by cryo-EM grid preparation, grid screening and preliminary data collection for single-particle analysis, or only the first two steps for cryo-ET. In both cases, if the screening steps are successfully completed, high-resolution data collection will be carried out using a Titan Krios microscope equipped with a latest-generation direct electron counting detector coupled to an energy filter. The SOS pipeline thus emerges as a comprehensive and efficient solution, further democratizing access to cryo-EM research.
摘要：
在应对有限（或没有）低温电子显微镜（cryo-EM）基础设施的实验室和大学所面临的挑战时，ESRF,与格勒诺布尔结构生物学研究所（IBS）合作，已经实施了cryo-EM解决方案到结构(SOS)管道。这个包容性的过程，从网格准备到高分辨率数据收集，涵盖单粒子分析和低温电子层析成像(cryo-ET)。可通过滚动访问路线访问，建议经过科学价值和技术可行性评估。严格的可行性标准需要样本同质性的有力证据。提供了两个不同的切入点：用户可以提交纯化的蛋白质样品进行全面处理，也可以使用已经玻璃化的低温EM网格启动管道。SOS管道集成了阴性染色成像（蛋白质样品专用）作为第一个质量步骤，然后是低温EM网格准备，用于单粒子分析的网格筛选和初步数据收集，或者只有冷冻ET的前两个步骤。在这两种情况下，如果筛选步骤成功完成，高分辨率数据收集将使用TitanKrios显微镜进行，该显微镜配备了最新一代的直接电子计数探测器和能量过滤器。因此，SOS管道成为一种全面高效的解决方案，进一步民主化的冷冻EM研究。
