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An All-in-one Sample Holder for Macromolecular X-ray Crystallography with Minimal Background Scattering
Published on: July 6, 2019
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A microfabricated fixed path length silicon sample holder improves background subtraction for cryoSAXS.
Jesse B Hopkins1, Andrea M Katz2, Steve P Meisburger2
1Laboratory of Atomic and Solid State Physics, Cornell University , Ithaca, New York 14853, USA.
Journal of Applied Crystallography
|June 20, 2015
Summary
Cryocooling biological samples for small-angle X-ray scattering (SAXS) significantly reduces radiation damage. Microfabricated silicon sample holders enable reproducible vitrified samples for faster, routine cryoSAXS data collection.
Area of Science:
- Biophysics
- Structural Biology
- Biochemistry
Background:
- Small-angle X-ray scattering (SAXS) is crucial for characterizing biological macromolecules in solution.
- Radiation damage and large sample volumes limit high-throughput SAXS applications.
- Cryocooling reduces radiation damage but requires identically vitrified samples for accurate background subtraction.
Purpose of the Study:
- To develop and validate microfabricated silicon sample holders for cryoSAXS.
- To overcome limitations in reproducible sample preparation for cryoSAXS.
- To enable faster and more routine high-throughput characterization of biomolecules.
Main Methods:
- Microfabrication of fixed path length silicon sample holders.
- Cryocooling of biological samples within the silicon holders.
- Small-angle X-ray scattering (SAXS) data collection and analysis.
Main Results:
- The silicon sample holders exhibit low background scattering and X-ray absorption.
- Only 640 nanoliters of sample are required, with reproducible cooling achieved.
- Data collected from 2.5 nanoliter illuminated volumes are reproducible down to q ≃ 0.02 Å⁻¹.
- CryoSAXS data quality is sufficient for reconstructions matching crystal structures.
Conclusions:
- Microfabricated silicon sample holders facilitate faster, more routine cryoSAXS data collection.
- These holders address challenges in reproducible sample vitrification and background subtraction.
- Further development is needed to minimize sample fracturing and enhance low-q data quality.
