4.2 Article

Solution scattering at the Life Science X-ray Scattering (LiX) beamline

Journal

JOURNAL OF SYNCHROTRON RADIATION
Volume 27, Issue -, Pages 804-812

Publisher

INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S1600577520002362

Keywords

SAXS; WAXS; solution scattering; SEC-SAXS; automation

Funding

  1. National Institute of Health, National Institute of General Medical Sciences [P41 GM111244]
  2. DOE Office of Biological and Environmental Research [KP1605010]
  3. NIH [S10 OD012331]
  4. US Department of Energy, Office of Science [DE-SC0012704]

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This work reports the instrumentation and software implementation at the Life Science X-ray Scattering (LiX) beamline at NSLS-II in support of biomolecular solution scattering. For automated static measurements, samples are stored in PCR tubes and grouped in 18-position sample holders. Unattended operations are enabled using a six-axis robot that exchanges sample holders between a storage box and a sample handler, transporting samples from the PCR tubes to the X-ray beam for scattering measurements. The storage box has a capacity of 20 sample holders. At full capacity, the measurements on all samples last for similar to 9 h. For in-line size-exclusion chromatography, the beamline-control software coordinates with a commercial high-performance liquid chromatography (HPLC) system to measure multiple samples in batch mode. The beamline can switch between static and HPLC measurements instantaneously. In all measurements, the scattering data span a wide q-range of typically 0.006-3.2 angstrom(-1). Functionalities in the Python package py4xs have been developed to support automated data processing, including azimuthal averaging, merging data from multiple detectors, buffer scattering subtraction, data storage in HDF5 format and exporting the final data in a three-column text format that is acceptable by most data analysis tools. These functionalities have been integrated into graphical user interfaces that run in Jupyter notebooks, with hooks for external data analysis software.

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