Smith Clyde A, Cohen Aina E
The Stanford Synchrotron Radiation Laboratory, Menlo Park, CA 94025, USA.
JALA Charlottesv Va. 2008 Dec 1;13(6):335-343. doi: 10.1016/j.jala.2008.08.008.
The macromolecular crystallography experiment lends itself perfectly to high-throughput technologies. The initial steps including the expression, purification and crystallization of protein crystals, along with some of the later steps involving data processing and structure determination have all been automated to the point where some of the last remaining bottlenecks in the process have been crystal mounting, crystal screening and data collection. At the Stanford Synchrotron Radiation Laboratory (SSRL), a National User Facility which provides extremely brilliant X-ray photon beams for use in materials science, environmental science and structural biology research, the incorporation of advanced robotics has enabled crystals to be screened in a true high-throughput fashion, thus dramatically accelerating the final steps. Up to 288 frozen crystals can be mounted by the beamline robot (the Stanford Automated Mounter, or SAM) and screened for diffraction quality in a matter of hours without intervention. The best quality crystals can then be remounted for the collection of complete X-ray diffraction data sets. Furthermore, the entire screening and data collection experiment can be controlled from the experimenter's home laboratory by means of advanced software tools that enable network-based control of the highly automated beamlines.
大分子晶体学实验非常适合高通量技术。包括蛋白质晶体的表达、纯化和结晶在内的初始步骤,以及一些涉及数据处理和结构测定的后续步骤,都已实现自动化,以至于该过程中最后剩下的一些瓶颈在于晶体安装、晶体筛选和数据收集。在斯坦福同步辐射实验室(SSRL),这是一个为材料科学、环境科学和结构生物学研究提供极其明亮的X射线光子束的国家用户设施,先进机器人技术的引入使得晶体能够以真正的高通量方式进行筛选,从而极大地加速了最后步骤。束线机器人(斯坦福自动安装仪,或SAM)可以在数小时内无需人工干预地安装多达288个冷冻晶体,并对其衍射质量进行筛选。然后可以重新安装质量最好的晶体以收集完整的X射线衍射数据集。此外,整个筛选和数据收集实验可以通过先进的软件工具在实验者的家庭实验室中进行控制,这些软件工具能够对高度自动化的束线进行基于网络的控制。