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来确定，包括x射线晶体学、核磁共振或电子显微镜。由于晶体学可以提供特定系统的一些最高分辨率视图，并且不受粒度的限制，因此这种方法的使用是广泛的，并为bbb领域贡献了最多的结构。x射线晶体学研究的一个固有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识别，这些条件对于给定的蛋白质、蛋白质复合物或蛋白质-核酸复合物是独特的。确定结晶条件涉及筛选目标，以对抗数百或数千种不同的化学物质，通常使用24或96孔托盘。理想情况下，托盘在无振动的环境中孵育，以帮助晶体形成，并在不同的温度下孵育（通常为4℃和18℃），以扩大搜索空间的深度。用光学显微镜定期观察托盘以寻找晶体的存在，这些晶体通常很小和/或不规则。获得这些初始“命中”的条件然后相互关联以寻找趋势，并优化以提高衍射研究的晶体质量。本提案中要求的Rigaku Minstrel成像系统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晶体托盘，并使用可见光和紫外光在预定的基础上自动拍摄每滴的数字图像。定期筛选允许随着时间的推移系统地覆盖结晶条件，大大增加了找到成功命中的可能性。紫外线照射可以发现隐藏在重沉淀物中的晶体，允许用户区分微晶体和颗粒沉淀物。紫外可视化进一步区分盐和蛋白质或核酸晶体，确保同步加速器源的宝贵x射线束时间有效地用于生物样品。托盘可以在4°C下孵育，由于缺乏冷室空间，我们无法达到这种状态，从而开辟了一个全新的低温筛选条件领域。此外，Minstrel系统将使用户能够更快、更彻底地覆盖结晶空间，加快对多种高度重要生物系统的结构调查。

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.