Department for Atomically Resolved Dynamics, Max-Planck-Institute for Structure and Dynamics of Matter, Luruper Chaussee 149, 22761 Hamburg, Germany.
Scientific Support Unit Machine Physics, Max-Planck-Institute for Structure and Dynamics of Matter, Luruper Chaussee 149, 22761 Hamburg, Germany.
J Synchrotron Radiat. 2020 Mar 1;27(Pt 2):360-370. doi: 10.1107/S1600577520000685. Epub 2020 Feb 27.
Serial synchrotron crystallography (SSX) is an emerging technique for static and time-resolved protein structure determination. Using specifically patterned silicon chips for sample delivery, the `hit-and-return' (HARE) protocol allows for efficient time-resolved data collection. The specific pattern of the crystal wells in the HARE chip provides direct access to many discrete time points. HARE chips allow for optical excitation as well as on-chip mixing for reaction initiation, making a large number of protein systems amenable to time-resolved studies. Loading of protein microcrystals onto the HARE chip is streamlined by a novel vacuum loading platform that allows fine-tuning of suction strength while maintaining a humid environment to prevent crystal dehydration. To enable the widespread use of time-resolved serial synchrotron crystallography (TR-SSX), detailed technical descriptions of a set of accessories that facilitate TR-SSX workflows are provided.
串联同步辐射晶体学（SSX）是一种新兴的静态和时间分辨蛋白质结构测定技术。使用专门设计的硅片进行样品输送，“命中-返回”（HARE）协议允许高效的时间分辨数据采集。HARE 芯片中晶体井的特定图案提供了对许多离散时间点的直接访问。HARE 芯片允许光学激发和片上混合以启动反应，使大量蛋白质系统适用于时间分辨研究。通过一种新颖的真空加载平台，可以将蛋白质微晶体加载到 HARE 芯片上，该平台可以在保持潮湿环境以防止晶体脱水的同时，精细调节吸力强度。为了实现时间分辨串联同步辐射晶体学（TR-SSX）的广泛应用，提供了一组便于 TR-SSX 工作流程的附件的详细技术说明。
