Minstrel HTUV Gallery 700 Automated Crystal Growth and Imaging System

描述(由申请人提供)：了解生物系统的分子机制需要功能和结构信息。生物分子的结构通常由几种方法中的一种确定，包括X射线结晶学、核磁共振或EM。由于结晶学可以提供特定体系的一些最高分辨率的视图，并且不受颗粒大小的限制，因此这种方法的使用非常广泛，并为该领域贡献了最多的结构[1]。X射线结晶学研究的一个内在挑战是对高质量晶体的要求。结晶依赖于能够支持生长的化学条件的鉴定，这些条件是特定蛋白质、蛋白质复合体或蛋白质-核酸复合体所特有的。确定结晶条件包括针对数百或数千种不同的化学物质筛选目标，通常使用24或96孔塔板。理想的情况是，托盘在无振动的环境中孵化以帮助晶体形成，并在不同的温度(通常为4？C和18？C)下孵化以扩大搜索空间的深度。光学显微镜定期观察托盘，以寻找晶体的存在，晶体通常很小和/或不规则。然后，对获得这些初始“命中”的条件进行交叉关联，以寻找趋势，并对其进行优化，以提高晶体质量以进行衍射研究。本提案中要求的Rigaku Minstrel成像系统提供的好处将改变加州大学伯克利分校进行结晶实验的方式。该仪器在给定的温度下培养水晶托盘，并使用可见光和紫外光按计划自动拍摄每一滴水的数字图像。定期筛查允许随着时间的推移系统地覆盖结晶条件，极大地增加了找到成功匹配的可能性。紫外光照射可以发现隐藏在沉淀物中的晶体，使用户能够区分微晶体和颗粒状沉淀物。UV可视化进一步区分盐和蛋白质或核酸晶体，确保同步加速器上宝贵的X射线束时间在生物样品上得到有效利用。托盘可以在4摄氏度下孵化，由于冷藏室空间不足，我们无法进入这种环境，从而为筛选打开了一个全新的低温条件领域。此外，Minstrel系统将使用户能够更快、更彻底地覆盖结晶空间，加快对各种高度重要的生物系统的结构研究。

DESCRIPTION (provided by applicant): Understanding the molecular mechanisms of biological systems requires both functional and structural information. Structures of biomolecules are typically determined by one of several methods, including x-ray crystallography, NMR, or EM. Because crystallography can afford some of the highest-resolution views of a particular system, and is not limited by the particle size, the use of this method is widespread, and has contributed the greatest numbers of structures to the field [1]. An innate challenge to x-ray crystallographic investigations is the requirement for high-quality crystals. Crystallization relies on the identification of chemical conditions that can support growth, which conditions are unique to a given protein, protein complex, or protein-nucleic acid complex. Identifying crystallization conditions involves screening a target against hundreds or thousands of different chemistries, typically using 24 or 96-well trays. Ideally, trays are incubated in a vibration-free environment to aid crystal formation, and are incubated at varied temperatures (usually 4¿C and 18¿C) to broaden the depth of the search space. Trays are viewed by light microscopy at regular intervals to search for the presence of crystals, which often are small and/or irregular. The conditions under which these initial "hits" are obtained are then cross-correlated to look for trends, and optimized to improve crystal quality for diffraction studies. The Rigaku Minstrel imaging system requested in this proposal offers benefits that will transform how crystallization experiments are conducted at UC Berkeley. The instrument incubates crystal trays at a given temperature and automatically takes digital images of each drop on a scheduled basis using both visible and UV light. Scheduled screening allows for systematic coverage of crystallization conditions over time, greatly increasing the likelihood of finding successful hits. UV illumination can uncover crystals hiding within heavy precipitates, allowing users to distinguish micro-crystals from granular precipitates. UV visualization further discriminates between salt and protein or nucleic acid crystals, ensuring that precious x-ray beam time at synchrotron sources is used efficiently on biological samples. Trays can be incubated at 4¿C, a regime inaccessible to us due to a paucity of cold-room space, thereby opening up an entirely new realm of low-temperature conditions for screening. In addition, the Minstrel system will enable users to more rapidly and thoroughly cover crystallization space, speeding structural investigations into a diverse number of highly significant biological systems.