Scanning structural mapping at the Life Science X-ray Scattering Beamline

Area of Science:

• Materials Science
• Biophysics
• Structural Biology

Background:

• Microbeam X-ray scattering requires precise instrumentation for detailed structural analysis.
• Existing methods faced limitations in data acquisition speed and sample-specific processing.
• The Life Science X-ray Scattering (LiX) beamline at NSLS-II sought to enhance its capabilities.

Purpose of the Study:

• To describe novel instrumentation and software for microbeam scattering and structural mapping.
• To enable adjustable beam sizes and simultaneous collection of small and wide-angle X-ray scattering (SAXS/WAXS) data.
• To develop a generalized data processing workflow for diverse biological samples.

Main Methods:

• Utilized a two-stage focusing scheme to achieve adjustable microbeam sizes (micrometers to millimeters).
• Employed multiple Pilatus detectors, including an in-vacuum Pilatus 900k detector for enhanced azimuthal coverage.
• Implemented fly scans and an in-vacuum sample environment to optimize data collection and minimize background scattering.

Main Results:

• Successfully generated microbeams and collected simultaneous SAXS/WAXS data with improved wide-angle coverage.
• Developed a generalized data processing workflow reducing data to reciprocal coordinates for user-friendly analysis.
• Demonstrated capabilities through in-vacuum mapping of onion epidermal cell walls and tomographic sectioning of poplar stems.

Conclusions:

• The new instrumentation and software significantly advance microbeam scattering and structural mapping capabilities at the LiX beamline.
• The generalized data processing workflow facilitates quantitative analysis and structural map construction for various samples.
• This work provides a powerful tool for investigating the structure of biological materials at the microscale.