High-density grids for efficient data collection from multiple crystals.
High-density grids for efficient data collection from multiple crystals.
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DOI：
10.1107/s2059798315020847
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发表时间：
2016-01
期刊：
影响因子：
--
通讯作者：
Cohen AE
中科院分区：
文献类型：
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作者：
Baxter EL;Aguila L;Alonso-Mori R;Barnes CO;Bonagura CA;Brehmer W;Brunger AT;Calero G;Caradoc-Davies TT;Chatterjee R;Degrado WF;Fraser JS;Ibrahim M;Kern J;Kobilka BK;Kruse AC;Larsson KM;Lemke HT;Lyubimov AY;Manglik A;McPhillips SE;Norgren E;Pang SS;Soltis SM;Song J;Thomaston J;Tsai Y;Weis WI;Woldeyes RA;Yachandra V;Yano J;Zouni A;Cohen AE
为了在同步加速器和XFEL源上进行高效的基于测角仪的样品输送，研制了一种高密度样品架。更高的吞吐量的方法来安装和收集数据，从多个小的和辐射敏感的晶体是重要的，以支持具有挑战性的结构调查，使用微焦点同步加速器光束线。此外，有效的样品输送方法对于在X射线自由电子激光（XFEL）源（如直线加速器相干光源（LCLS））上进行生产性飞秒晶体学实验至关重要。为了满足这些需求，已开发出可用作晶体生长和衍射数据收集的支架的高密度样品网格，并用于在同步加速器和XFEL源处进行基于角度计的高效样品递送。单个网格包含75个安装端口，并适合SSRL盒或单盘存储容器内。使用带有SSRL试剂卡的滤线栅将试剂卡容量扩展至7200份样本。网格也可以覆盖有聚合物薄膜或套筒，用于从多个样品中有效地收集室温数据。Blu-Ice/DCSS实验控制系统已纳入新的自动化程序，以支持网格，包括半自动网格对齐，网格端口的全自动定位，光栅化和自动数据收集。已经开发了专门的工具来支持网格上的结晶实验，包括通用适配器，该适配器允许由商业液体处理机器人填充网格，以及培养室，该培养室支持蒸气扩散和立方相结晶实验。描述了使用液体处理机器人和孵育室将晶体装载到网格中或在网格上生长的实验。在LCLS-XPP和SSRL BL 12 -2中在室温和低温下筛选晶体。
A high-density sample mount has been developed for efficient goniometer-based sample delivery at synchrotron and XFEL sources. Higher throughput methods to mount and collect data from multiple small and radiation-sensitive crystals are important to support challenging structural investigations using microfocus synchrotron beamlines. Furthermore, efficient sample-delivery methods are essential to carry out productive femtosecond crystallography experiments at X-ray free-electron laser (XFEL) sources such as the Linac Coherent Light Source (LCLS). To address these needs, a high-density sample grid useful as a scaffold for both crystal growth and diffraction data collection has been developed and utilized for efficient goniometer-based sample delivery at synchrotron and XFEL sources. A single grid contains 75 mounting ports and fits inside an SSRL cassette or uni-puck storage container. The use of grids with an SSRL cassette expands the cassette capacity up to 7200 samples. Grids may also be covered with a polymer film or sleeve for efficient room-temperature data collection from multiple samples. New automated routines have been incorporated into the Blu-Ice/DCSS experimental control system to support grids, including semi-automated grid alignment, fully automated positioning of grid ports, rastering and automated data collection. Specialized tools have been developed to support crystallization experiments on grids, including a universal adaptor, which allows grids to be filled by commercial liquid-handling robots, as well as incubation chambers, which support vapor-diffusion and lipidic cubic phase crystallization experiments. Experiments in which crystals were loaded into grids or grown on grids using liquid-handling robots and incubation chambers are described. Crystals were screened at LCLS-XPP and SSRL BL12-2 at room temperature and cryogenic temperatures.