Minstrel HTUV Gallery 700 Automated Crystal Growth and Imaging System
Minstrel HTUV Gallery 700 自动晶体生长和成像系统
基本信息
- 批准号:8447984
- 负责人:
- 金额:$ 37.66万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2013
- 资助国家:美国
- 起止时间:2013-06-01 至 2015-05-31
- 项目状态:已结题
- 来源:
- 关键词:BiologicalChemicalsChemistryComplexCrystal FormationCrystallizationCrystallographyDropsEnsureEnvironmentGrowthImageImageryIncubatedInvestigationLightingMethodsMolecularNucleic AcidsParticle SizeProteinsResolutionSamplingScheduleSodium ChlorideSourceSpeedStructureSynchrotronsSystemTemperatureTimeUltraviolet RaysVisible Radiationbasebiological systemscold temperaturedigital imagingimprovedinstrumentlight microscopyprotein complexresearch studyscreeningtrendvibration
项目摘要
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.
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通过多种方法之一来确定,包括 X 射线晶体学、NMR 或 EM,因为晶体学可以提供一些信息。特定系统的最高分辨率视图,并且不受颗粒尺寸的限制,这种方法的使用非常广泛,并且为该领域贡献了最大数量的结构[1]。晶体学研究是高质量晶体的要求,结晶依赖于支持生长的化学条件的鉴定,这些条件对于给定的蛋白质、蛋白质复合物或蛋白质-核酸复合物来说是独特的,鉴定结晶条件涉及筛选目标。针对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,托盘在无振动环境中孵育以帮助晶体形成,并在不同温度(通常为 4°C)下孵育。 C 和 18¿ C) 扩大搜索空间的深度 通过光学显微镜定期观察托盘,以搜索晶体的存在,这些晶体通常很小和/或不规则。然后获得这些初始“命中”的条件。该提案中要求的 Rigaku Minstrel 成像系统将改变加州大学伯克利分校结晶实验的进行方式。指定的温度,并使用可见光和紫外光自动拍摄每个液滴的数字图像。预定的筛选可以随着时间的推移系统地覆盖结晶条件,大大增加了成功发现隐藏在重物中的晶体的可能性。沉淀物,使用户能够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。托盘可在 4℃ 下孵育C,这是由于冷室空间不足而无法进入的区域,从而开辟了一个全新的低温条件筛选领域。此外,Minstrel 系统将使用户能够更快速、更彻底地覆盖结晶空间,加快对多种高度重要的生物系统的结构研究。
项目成果
期刊论文数量(1)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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