Glacios cryo-electron microscope for single particle analysis and electron tomography of proteins, complexes and fibrillar assemblies

几十年来，电子显微镜一直被用来提供细胞和组织结构的细节，并对光学显微镜无法看到的颗粒(如病毒)进行成像。然而，近年来，随着冷冻样品技术的改进以及软件和硬件的进步，电子显微镜经历了一场“分辨率革命”。现在可以从CryoEM数据中观察到的细节包括蛋白质的结构到原子分辨率，即可以看到分子中所有原子的水平。近年来，“拆分革命”导致了许多新的结构被确定，并因这些进展而在2017年诺贝尔化学奖上获得认可。重要的是，以前很难确定其原子分辨率结构的蛋白质现在第一次使用低温EM数据进行成像。在曼彻斯特大学，我们有许多由BBSRC资金支持的项目，需要访问低温EM。然而，我们目前的显微镜Polara已经过时，不再受到制造商的支持。因此，这项应用是为了提供最先进的冰川冷冻电子显微镜，以支持曼彻斯特大学一个非常富有成效的科学家团队的研究。由于自动化的进步，冰川冷冻EM是显微镜技术的一大进步，它将使我们能够以更快的速度收集数据，这意味着每天可以收集更多的图像。冰川也更加稳定，这将使我们能够收集更高分辨率的图像。此外，冰川具有与其他仪器兼容的低温EM样本处理，包括国家电子生物成像中心的仪器，这意味着样本可以在显微镜之间转移以收集数据。曼彻斯特的研究需要获得新的低温EM，涵盖了与BBSRC战略重点一致的广泛科学。项目包括分析参与细胞信号、组织强度和炎症的细胞外基质蛋白；确定导致多药耐药和肾脏功能的膜蛋白的结构；研究生物催化的酶机制；了解合成蛋白质所涉及的过程；以及在新疫苗的开发中分析类似病毒的颗粒。这些研究将极大地受益于新的低温EM，曼彻斯特大学认识到这一点，提供了主要的财政支持，包括冰川成本的35%，以及我们电子显微镜设施中专门建造的房间和一名将操作显微镜和培训新用户的专家实验官员。

Electron microscopy has been used for decades to provide details of cell and tissue structures and to image particles that could not be seen by light microscopes, such as viruses. However, in recent years electron microscopy has undergone a "Resolution Revolution" with a combination of improvements in freezing samples (cryoEM) along with software and hardware advances. The details that can now be observed from cryoEM data include the structure of proteins to atomic resolution, the level where all atoms in the molecule can be seen. The "Resolution Revolution" has led to many new structures being determined in recent years and was acknowledged in the award of the 2017 Nobel Prize in Chemistry for these developments. Importantly, proteins that were previously too difficult to determine their atomic resolution structure are now being imaged for the first time using cryoEM data.At the University of Manchester, we have many projects supported by BBSRC funding that require access to cryoEM. However, our current microscope, the Polara, is out-dated and no longer supported by the manufacturer. Therefore, this application is to provide a state-of-the-art Glacios cryo-electron microscope to support the research of a very productive team of scientists at the University of Manchester. The Glacios cryoEM is a step-change in microscope technology which would allow us to collect data at a much faster rate, due to advances in automation, which means more images can be collected each day. The Glacios is also much more stable which will allow us to collect higher resolution images. In addition, the Glacios has compatible cryoEM sample handling with other instruments, including those in the national electron bioimaging centre, which means samples can be transferred between microscopes for data collection.The research at Manchester that needs access to a new cryoEM covers a broad range of science that is in alignment with BBSRC's strategic priorities. Projects include the analysis of extracellular matrix proteins involved in cell signalling, tissue strength and inflammation; determining the structures of membrane proteins responsible for multidrug resistance and kidney function; investigation of enzyme mechanism for biological catalysis; understanding the processes involved in synthesising proteins, and analysing virus-like particles in the development of new vaccines. These studies will benefit enormously from a new cryoEM and the University of Manchester recognises this by offering major financial support including 35% of the cost of the Glacios, plus a purpose-built room in our Electron Microscopy Facility and an expert experimental officer who will operate the microscope and train new users.