A cryo-capable electron microscope for the Astbury Biostructure Laboratory

结构生物学是生物学发现的有力工具。它可以揭示蛋白质和其他分子如何相互作用的分子细节，从而帮助我们理解在人类/动物健康和福祉、食品安全和生物技术等不同领域具有巨大影响的基本生物过程。现代结构生物学的基石之一是一种被称为电子显微镜的技术，在这种技术中，强大的显微镜被用来拍摄生物材料（如蛋白质）的高度放大图像，从中我们可以计算出蛋白质复合物的三维结构。冷冻样品成像技术（CryoEM）现在可以解决前所未有的规模和复杂性的结构问题，其结果正在改变我们对生物学的理解。利兹大学的阿斯特伯里生物结构实验室是一家国际公认的冷冻设备，为来自英国和其他国家的大学和工业界的大量科学家提供支持。我们目前使用两台刚刚更新到最新技术的超高端电子显微镜。然而，当新的样品到达我们的工厂，新的项目开始时，这些高端显微镜处于低端电子显微镜管道的末端。对于每一个这样的样品，常规的电子显微镜被用来快速成像样品，并评估其质量和高分辨率成像的适用性。常规电子显微镜对于优化样品制备高端成像的过程也是绝对必要的。因此，常规成像对于最终导致高分辨率结构的每个项目都是必不可少的。然而，我们在阿斯特伯里生物结构实验室用于常规成像的显微镜（以及显微镜上的探测器）正在迅速过时，变得难以维护和昂贵，而且不经济或不可能升级。因此，我们希望从这项拨款申请中获得资金，购买一台新的电子显微镜，配备现代化的探测器，以及允许远程操作的现代软件，用于常规成像和样品优化。这将使我们能够淘汰两台老化的显微镜，减少我们的空间和成本，并减少我们设施的碳足迹。新的显微镜将支持目前正在进行的所有令人兴奋的结构项目，该应用程序描述了四个主要主题中的一个子集：(1)位于细胞周围保护膜上的蛋白质；(2)病毒的结构和功能；(3)大的蛋白质组合；(4)聚集的蛋白质。此外，Astbury生物结构实验室现在是冷冻电镜培训的主要中心，新的显微镜将直接支持这一活动，帮助我们提高使用冷冻电镜进行生物发现所需的技能。

Structural biology is a powerful tool in biological discovery. It can reveal the molecular details of how proteins and other molecules interact, and so help us understand fundamental biological processes that have enormous implications in areas as diverse as human/animal health and wellbeing, food security and biotechnology. One of the cornerstones of modern structural biology is a technique called electron microscopy, where powerful microscopes are used to take highly magnified images of biological material such as proteins, from whcih we can work out the three-dimensional structure of the protein complexes. CryoEM (imaging frozen samples) can now address structural problems of unprecedented scale and complexity, and the results are transforming our understanding of biology.The Astbury Biostructure Laboratory at the University of Leeds is an internationally-recognised cryoEM facility that supports large numbers of scientists from Universities and Industry, and from the UK and beyond. We currently operate two ultra-high-end electron microscopes that have just been updated to the latest technology. However when new samples arrive in our facility, and new projects are started, these high-end microscopes sit at the end of a pipeline that starts with lower-end electron microscopes. For every such sample, routine electron microscopes are used to quickly image a sample and assess its quality and suitability for high-resolution imaging. Routine electron microscopes are also absolutely essential to the process of optimising how samples are prepared for high end-imaging. Routine imaging is therefore essential to every project that ultimately leads to a high resolution structures. However, the microscopes available to us for routine imaging at the Astbury Biostructure Laboratory (and the detectors on those microscopes) are rapidly becoming obsolete, becoming difficult and expensive to maintain, and uneconomic or impossible to upgrade. We therefore wish to secure funding from this grant application to buy a new electron microscope, with a modern detector, and modern software allowing remote operation, for routine imaging and sample optimisation. This will allow us to retire two ageing microscopes, reducing our space and costs, and shrinking the carbon footprint of our facility. The new microscope will support all of the exciting structural projects currently underway, and this application describes a subset of these in four major themes: (1) proteins that sit in the protect membranes around cells, (2) the structure and function of viruses; (3) large protein assemblies; and (4) proteins that aggregate. In addition, the Astbury Biostructure Laboratory is now a major centre for cryoEM training, and the new microscope will directly support this activity, helping us to drive an increase in the skills needed to use cryoEM for biological discovery.