Acquisition of Automated Nanoscale Crystallization Equipment

描述（由申请人提供）：在此提案中，麻省理工学院生物系和化学系的六名教员请求资助在麻省理工学院结构生物学核心设施安装自动化大分子结晶设备。直到今年，麻省理工学院的结构生物学家还缺乏在大分子晶体学设施中日益常见的自动化纳米级结晶设备。结晶筛选和优化是通过手动移液进行的。随后的观察也是手动的。这种实验方法费力、重复且浪费——消耗大量生产成本昂贵的样品。在结构蛋白质组学项目的推动下，最近的技术发展已经提供了可用的设备，这些设备现在在筛选和初始优化方面很大程度上取代了手动方法。作为第一步，这组结构生物学家最近购买了用于纳米级结晶筛选的自动液体处理系统。前两个月的结果超出了预期：样本材料有限的项目已成为可能并已启动，提高的可重复性启用了重点优化策略，并在创纪录的时间内找到了初步命中。所申请的资金将用于扩展和补充该液体处理系统，为最新的结晶设施提供必要的设备，即组成精确定制的网格屏幕的移液机器人、用于跟踪结晶试验的组合成像和存储系统，以及具有足够容量的存储和网络服务器，用于记录并向研究人员呈现结晶实验的预定成像结果。这个集成系统将极大地促进麻省理工学院结构生物学的发展。它将允许学生和博士后科学家根据当今的标准进行系统的结晶实验。它将使得纯化样品量受到限制的项目变得可行——所用蛋白质减少 10-15 倍是一个关键优势，因为我们的许多目标是需要在真核细胞中表达的大分子复合物或蛋白质。最后，该系统将节省其他方面的资金，包括花费的带薪时间和消耗的消耗品，特别是在困难的大分子晶体学项目上。公共卫生相关性：主要用户的研究兴趣涵盖现代生物学和生物化学的广度。拟议的设备将使我们能够更系统、更高效地开展结构项目。它将使我们能够追求当前生物学和生物医学领域最感兴趣的结构目标，而巧合的是，这些结构目标通常是最难生产足够的高度纯化的样品材料的目标。

DESCRIPTION (provided by applicant): In this proposal, six faculty of the Departments of Biology and of Chemistry at MIT request funding for automated macromolecular crystallization equipment to be installed at MIT's Structural Biology Core Facility. MIT structural biologists have lacked, until this year, any of the automated, nanoscale crystallization equipment that is becoming common at macromolecular crystallography facilities. Crystallization screens and optimization have been performed by manually pipetting. Subsequent observation is manual as well. This experimental approach is laborious, repetitive, and wasteful - consuming large amounts of samples that are expensive to produce. Recent technology developments, driven by structural proteomics projects, have made available equipment that now largely supplants manual methods in screening and initial optimization. As a first step, this group of structural biologists recently purchased an automated liquid handling system for nanoscale crystallization screening. The results in the first two months have exceeded expectations: projects where sample material was limiting have become possible and have been initiated, improved reproducibility has enabled focused optimization strategies and initial hits have been found in record time. The requested funds will be used to expand and complement this liquid handling system, providing the equipment necessary to an up-to-date crystallization facility - namely, a pipetting robot to compose accurately customized grid screens, a combined imaging and storage system to follow crystallization trials, and a storage and web server with sufficient capacity to record and present to investigators the results of scheduled imaging of crystallization experiments. This integrated system will promote immeasurably structural biology at MIT. It will allow students and postdoctoral scientists to perform systematically crystallization experiments according to today<s standards. It will make feasible projects for which the amount of purified sample is limiting -- 10-15 fold reduction in protein used is a key benefit, because many of our targets are macromolecular complexes or proteins requiring expression in eukaryotic cells. Lastly, the system will save funds elsewhere, in salaried time spent and consumables expended, especially on difficult macromolecular crystallography projects. PUBLIC HEALTH RELEVANCE: The research interests of the primary users span the breadth of modern biology and biochemistry. The proposed equipment will allow us to pursue structural projects more systematically and thus more efficiently. It will enable us to pursue those structural targets of the most current biological and biomedical interest, which are, coincidentally, often the ones for which sufficient highly purified sample material is the hardest to produce.