BioSAXS on the SWING beamline at Synchrotron SOLEIL
BioSAXS on the SWING beamline at Synchrotron SOLEIL
复制标题
Synchrotron SOLEIL 的 SWING 光束线上的 BioSAXS
DOI：
10.1107/s1600576721008736
复制
发表时间：
2021
影响因子：
6.1
通讯作者：
Javier Pérez
中科院分区：
文献类型：
--
作者：
A. Thureau;P. Roblin;Javier Pérez
溶液中蛋白质的小角X射线散射（SAXS）已成为生物化学家和结构生物学家的重要工具，特别是由于同步辐射源具有高通量能力的光束线的可用性。尽管自2008年开放以来，SWING光束线处理了大量的科学学科，但一直致力于为科学界提供最先进的生物SAXS（BioSAXS）仪器和数据简化方法。极其可靠的真空EigerX-4 M探测器允许无噪声地收集无限数量的帧。一个小的光束停止包括金刚石二极管为基础的监视器，使测量的透射强度与0.1%的精度，以及一个q
最大/q
在单个距离处最小比率高达140。安装新的混合叶片大大减少了寄生散射。一种新的热控制的真空毛细管保持器，包括光纤为基础的光谱功能，允许同时使用三种光谱技术，除了SAXS测量。添加第二个高效液相色谱（HPLC）回路实际上消除了与柱平衡相关的等待时间。多角度光散射光谱仪和折射仪的简单在线连接允许分别独立测定低紫外吸收样品（如洗涤剂和糖）的分子量和浓度。这些工具性的改进与重要的软件开发相结合。HPLC进样Agilent软件由SAXS光束线采集软件控制，允许几乎无限系列的自动SAXS测量与样品进样同步。所有包含数据的文件和报告都自动存储在相同的文件夹中，其名称与用户和样品相关。此外，所有SAXS原始图像都在运行中自动处理，分析数据存储在ISPyB数据库中，并可通过网页访问。
Small-angle X-ray scattering (SAXS) of proteins in solution has become a key tool for biochemists and structural biologists, thanks especially to the availability of beamlines with high-throughput capabilities at synchrotron sources. Despite the large spectrum of scientific disciplines tackled on the SWING beamline since its opening in 2008, there has always been a strong commitment to offering state-of-the-art biological SAXS (BioSAXS) instrumentation and data reduction methods to the scientific community. The extremely reliable in-vacuum EigerX-4M detector allows collection of an unlimited number of frames without noise. A small beamstop including a diamond diode-based monitor enables measurements of the transmitted intensity with 0.1% precision as well as a q
max/q
min ratio as large as 140 at a single distance. The parasitic scattering has been strongly reduced by the installation of new hybrid blades. A new thermally controlled in-vacuum capillary holder including fibre-optics-based spectroscopic functionalities allows the simultaneous use of three spectroscopic techniques in addition to SAXS measurements. The addition of a second high-performance liquid chromatography (HPLC) circuit has virtually eliminated the waiting time associated with column equilibration. The easy in-line connection of a multi-angle light scattering spectrometer and a refractometer allows for an independent determination of the molecular mass and of the concentration of low-UV-absorption samples such as detergents and sugars, respectively. These instrumental improvements are combined with important software developments. The HPLC injection Agilent software is controlled by the SAXS beamline acquisition software, allowing a virtually unlimited series of automated SAXS measurements to be synchronized with the sample injections. All data-containing files and reports are automatically stored in the same folders, with names related to both the user and sample. In addition, all raw SAXS images are processed automatically on the fly, and the analysed data are stored in the ISPyB database and made accessible via a web page.