ROBOTIC CRYSTAL MOUNTING & ALIGNMENT SYSTEMS

机器人晶体安装

• 批准号: 6652979
• 负责人: Thomas N Earnest
• 金额: $ 1.01万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6652979
• 关键词: X ray crystallography; bioengineering /biomedical engineering; bioimaging /biomedical imaging; biomedical equipment development; computer program /software; cryoscience; crystallization; data collection methodology /evaluation; fiber optics; fluorescence spectrometry; high throughput technology; imaging /visualization /scanning; micromanipulator; protein structure; proteomics; robotics; structural biology

This proposal is to develop and test an integrated assembly of robotics subsystems dedicated to high-throughput protein crystallography at a synchrotron facility. The long-term goal is the implementation of an automated environment for the efficient manipulation of large numbers of single crystal protein samples for x-ray diffraction intensity collection at synchrotron-based biological crystallography facilities. Specific subtasks include the design and implementation of manual, semi- automated, and automated remote control systems for sample manipulation on the goniometer and in x-ray beam, development of algorithms for automated centering of target features, design and testing of a subsystem to load and unload conventional cryogenically-frozen protein crystal samples, development of protocols for screening large numbers of crystals for data collection, development and testing of new sample holders for efficient crystal harvesting, storage, manipulation, and data collection, and finally, adaptation of the hardware to the alignment of microcrystals. Although there are currently more than a dozen synchrotron beamlines in the U.S. dedicated to protein crystallography, the throughput is limited by the speed of manual manipulation (loading, centering, and unloading). At the Macromolecular Crystallography Facility at Advanced Light Source, one current beamline, two new beamlines under construction, and several new beamlines being designed, will benefit from the use of automated stages -initially a simple x,y,z movement with motors under manual control - for centering protein crystals. The proposed integrated approach for automated manipulation of protein crystals will be more efficient (in terms of time and synchrotron floor space), and will substantially increase throughput and reliably in handling samples. This proposal is in direct response to the recommendations of the NIH and DOE in designing automated systems for use at synchrotron facilities and in developing technologies necessary for advancing high-throughput structural genomics initiatives.

该建议是开发和测试一个集成的组装机器人子系统，致力于高通量蛋白质晶体在同步加速器设施。长期目标是实现一个自动化的环境，用于在基于同步加速器的生物晶体学设施中有效操作大量单晶蛋白质样品，以进行X射线衍射强度收集。具体的子任务包括设计和实现用于在测角仪上和在X射线束中进行样品操作的手动、半自动和自动远程控制系统，开发用于目标特征的自动定心的算法，设计和测试用于装载和卸载常规低温冷冻蛋白质晶体样品的子系统，开发用于筛选大量晶体以用于数据收集的协议，开发和测试新的样品架，用于有效的晶体收获、存储、操作和数据收集，最后，使硬件适应微晶的排列。虽然目前在美国有十几条同步加速器光束线专门用于蛋白质晶体学，但产量受到手动操作（加载，定心和卸载）速度的限制。在Advanced Light Source的高分子晶体学设施中，一条现有的光束线，两条正在建设中的新光束线和几条正在设计的新光束线将受益于自动化平台的使用-最初是一个简单的x，y，z运动，手动控制电机-用于定心蛋白质晶体。所提出的用于蛋白质晶体自动化操作的集成方法将更有效（在时间和同步加速器占地面积方面），并且将大幅提高处理样品的吞吐量和可靠性。该提案是对NIH和DOE在设计用于同步加速器设施的自动化系统和开发推进高通量结构基因组学计划所需技术方面的建议的直接回应。