ROBOTIC CRYSTAL MOUNTING & ALIGNMENT SYSTEMS

这项提议是在同步加速器设施中开发和测试专门用于高通量蛋白质结晶学的机器人子系统的集成组装。长期目标是实现一个自动化环境，以便在基于同步加速器的生物结晶学设施中有效地处理大量单晶蛋白质样品，以便收集X射线衍射强度。特定子任务包括设计和实施手动、半自动和自动遥控系统，用于在测角仪和X射线束中操纵样品，开发用于目标特征自动居中的算法，设计和测试用于加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用于数据收集的协议，开发和测试用于高效晶体采集、存储、操纵和数据收集的新样品保持器，以及最后，使硬件适应微晶体的排列。尽管目前美国有十几条同步加速器光束线专门用于蛋白质结晶学，但吞吐量受到手动操作(加载、对中和卸载)速度的限制。在先进光源的高分子结晶设施，一条当前的光束线，两条在建的新光束线，以及几条正在设计的新光束线，将受益于自动工作台的使用-最初是简单的x，y，z移动，在手动控制下的电机-居中蛋白质晶体。建议的蛋白质晶体自动化操作的综合方法将更高效(在时间和同步加速器占地面积方面)，并将显著增加处理样品的吞吐量和可靠性。这项提议是对美国国立卫生研究院和美国能源部在设计用于同步加速器设施的自动化系统以及开发推进高通量结构基因组学倡议所需技术方面的建议的直接回应。

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.