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

几十年来，电子显微镜一直用于提供细胞和组织结构的细节，并为光学显微镜无法看到的颗粒（如病毒）成像。然而，近年来电子显微镜经历了一场“分辨率革命”，结合了冷冻样品（cryoEM）的改进以及软件和硬件的进步。现在可以从冷冻电镜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，“分辨率革命”导致了许多新结构的确定，并因此获得了2017年诺贝尔化学奖。重要的是，以前难以确定其原子分辨率结构的蛋白质现在首次使用低温电子显微镜数据进行成像。在曼彻斯特大学，我们有许多由BBSRC资助的项目需要使用cryoEM。然而，我们目前的显微镜，Polara，是过时的，不再支持的制造商。因此，该应用程序将提供最先进的Glacios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。Glacios cryoEM是显微镜技术的一个阶段性变化，由于自动化的进步，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。Glacios也更加稳定，这将使我们能够收集到更高分辨率的图像。此外，Glacios具有与其他仪器（包括国家电子生物成像中心的仪器）兼容的冷冻电镜样品处理功能，这意味着样品可以在显微镜之间转移以进行数据收集。曼彻斯特大学的研究需要获得一种新的冷冻电镜，涵盖了广泛的科学领域，与BBSRC的战略重点保持一致。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；确定与多药耐药和肾功能有关的膜蛋白结构；酶对生物催化作用机理的研究了解合成蛋白质的过程，并分析新疫苗开发过程中的病毒样颗粒。这些研究将从新的冷冻电镜中受益匪浅，曼彻斯特大学认识到这一点，提供了主要的财政支持，包括35%的Glacios成本，加上我们的电子显微镜设施的专用房间和一位专家实验官员，他将操作显微镜并培训新用户。

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.