Versatile sample environments and automation for biological solution X-ray scattering experiments at the P12 beamline (PETRA III, DESY).
Versatile sample environments and automation for biological solution X-ray scattering experiments at the P12 beamline (PETRA III, DESY).
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DOI：
10.1107/s160057671500254x
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发表时间：
2015-04-01
影响因子：
6.1
通讯作者：
Svergun DI
中科院分区：
文献类型：
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作者：
Blanchet CE;Spilotros A;Schwemmer F;Graewert MA;Kikhney A;Jeffries CM;Franke D;Mark D;Zengerle R;Cipriani F;Fiedler S;Roessle M;Svergun DI
在EMBL（DESY，汉堡）的高亮度P12同步加速器光束线上的生物小角X射线散射（BioSAXS）的综合环境允许广泛的溶液散射实验。设计了自动化硬件和软件系统，以确保数据收集和处理高效、精简和方便用户。位于PETRA III储存环（DESY，汉堡）的EMBL的高亮度同步加速器P12光束线专用于生物小角X射线散射（SAXS），并已被设计和优化用于大分子溶液的散射实验。无散射狭缝减少了寄生散射，定制设计的微型有源光束光阑确保了精确的数据归一化，光子计数PILLENT 2 M探测器实现了弱散射信号的无背景检测。高通量和小光束尺寸允许曝光时间低至30-50 ms的快速实验，覆盖约300至0.5 nm的分辨率范围。 P12拥有多功能和灵活的样品环境系统，可满足研究大分子溶液所需的各种实验需求。其中包括用于标准批次样品分析的真空毛细管模式，以及机器人样品输送和连续流动在线样品纯化和表征，以及空气中毛细管时间分辨停流设置。一种新型的微流控离心混合装置（SAXS盘）开发的高通量筛选模式，使用亚微升的样品体积。自动化是P12的一个关键功能;它由一个光束线Meta服务器控制，该服务器协调和安排标准或非标准操作设置的实验。集成的SASFLOW管道自动检查一致性，处理和分析数据，提供对整体参数的近实时评估，并在数据收集后几分钟内生成低分辨率模型。这些进步，结合远程访问选项，允许快速高通量分析，以及时间分辨和筛选实验的新手和专家生物SAXS用户。
An integrated environment for biological small-angle X-ray scattering (BioSAXS) at the high-brilliance P12 synchrotron beamline of the EMBL (DESY, Hamburg) allows for a broad range of solution scattering experiments. Automated hardware and software systems have been designed to ensure that data collection and processing are efficient, streamlined and user friendly. A high-brilliance synchrotron P12 beamline of the EMBL located at the PETRA III storage ring (DESY, Hamburg) is dedicated to biological small-angle X-ray scattering (SAXS) and has been designed and optimized for scattering experiments on macromolecular solutions. Scatterless slits reduce the parasitic scattering, a custom-designed miniature active beamstop ensures accurate data normalization and the photon-counting PILATUS 2M detector enables the background-free detection of weak scattering signals. The high flux and small beam size allow for rapid experiments with exposure time down to 30–50 ms covering the resolution range from about 300 to 0.5 nm. P12 possesses a versatile and flexible sample environment system that caters for the diverse experimental needs required to study macromolecular solutions. These include an in-vacuum capillary mode for standard batch sample analyses with robotic sample delivery and for continuous-flow in-line sample purification and characterization, as well as an in-air capillary time-resolved stopped-flow setup. A novel microfluidic centrifugal mixing device (SAXS disc) is developed for a high-throughput screening mode using sub-microlitre sample volumes. Automation is a key feature of P12; it is controlled by a beamline meta server, which coordinates and schedules experiments from either standard or nonstandard operational setups. The integrated SASFLOW pipeline automatically checks for consistency, and processes and analyses the data, providing near real-time assessments of overall parameters and the generation of low-resolution models within minutes of data collection. These advances, combined with a remote access option, allow for rapid high-throughput analysis, as well as time-resolved and screening experiments for novice and expert biological SAXS users.