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首页> 外文期刊>Journal of synchrotron radiation >Multi-channel in situ dynamic light scattering instrumentation enhancing biological small-angle X-ray scattering experiments at the PETRA III beamline P12
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Multi-channel in situ dynamic light scattering instrumentation enhancing biological small-angle X-ray scattering experiments at the PETRA III beamline P12

机译：MOVER-CHANDIN III X射线散射仪在PETRA III梁线P12中提高生物小角X射线散射实验

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摘要

Small-angle X-ray scattering (SAXS) analysis of biomolecules is increasingly common with a constantly high demand for comprehensive and efficient sample quality control prior to SAXS experiments. As monodisperse sample suspensions are desirable for SAXS experiments, latest dynamic light scattering (DLS) techniques are most suited to obtain non-invasive and rapid information about the particle size distribution of molecules in solution. A multi-receiver four-channel DLS system was designed and adapted at the BioSAXS endstation of the EMBL beamline P12 at PETRA III (DESY, Hamburg, Germany). The system allows the collection of DLS data within round-shaped sample capillaries used at beamline P12. Data obtained provide information about the hydrodynamic radius of biological particles in solution and dispersity of the solution. DLS data can be collected directly prior to and during an X-ray exposure. To match the short X-ray exposure times of around 1 s for 20 exposures at P12, the DLS data collection periods that have been used up to now of 20 s or commonly more were substantially reduced, using a novel multichannel approach collecting DLS data sets in the SAXS sample capillary at four different neighbouring sample volume positions in parallel. The setup allows online scoring of sample solutions applied for SAXS experiments, supports SAXS data evaluation and for example indicates local inhomogeneities in a sample solution in a time-efficient manner. Biological macromolecules with different molecular weights were applied to test the system and obtain information about the performance. All measured hydrodynamic radii are in good agreement with DLS results obtained by employing a standard cuvette instrument. Moreover, applying the new multi-channel DLS setup, a reliable radius determination of sample solutions in flow, at flow rates normally used for size-exclusion chromatography-SAXS experiments, and at higher flow rates, was verified as well. This study also shows and con
机译：小角X射线散射（SAX）生物分子的分析越来越常见，在萨克斯实验之前对全面和高效的样品质量控制的不断需求越来越多。由于单分散样品悬浮液对于SAXS实验期望，最新的动态光散射（DLS）技术最适合于获得关于溶液中分子粒度分布的非侵入性和快速信息。设计了一种多接收器四通道DLS系统，并适用于Petra III（德国斯花，德国）的Embl BeamLine P12的Biosaxs封端。该系统允许在光束线P12使用的圆形样品毛细管内收集DLS数据。获得的数据提供有关溶液溶液中的生物颗粒的流体动力半径的信息和溶液的分散性。 DLS数据可以在X射线曝光之前和期间直接收集。为了匹配在P12的20个曝光的短X射线曝光时间为20个曝光，使用新颖的多通道方法收集DLS数据集的新型多通道方法，已经使用的DLS数据收集周期为20 s或通常更多地减少在SAXS样品中，在四个不同相邻的样本体积的样本中并联位置。该设置允许施加用于萨克斯实验的样本溶液的在线评分，支持萨克斯数据评估，例如，以较效的方式表示样品溶液中的局部不均匀性。应用具有不同分子量的生物大分子以测试系统并获得有关性能的信息。所有测量的流体动力学半径都与通过采用标准比色皿仪器获得的DLS结果一致。此外，还验证了应用新的多通道DLS设置，以通常用于尺寸排除色谱 - 撒酷淋巴实验的流量的可靠半径确定样品溶液的样品溶液。这项研究还显示和骗局

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