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Microcrystallography of Protein Crystals and In Cellulo Diffraction
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Detector alignment for X-ray crystallography using Millepede-II.

Thomas A White1,2

  • 1Deutsches Elektronen-Synchrotron DESY Hamburg Germany.

Journal of Applied Crystallography
|April 10, 2026
PubMed
Summary
This summary is machine-generated.

A new method refines X-ray detector geometry using the Millepede algorithm, improving serial crystallography data accuracy. This approach efficiently corrects detector parameters, enhancing crystal structure determination and data quality in experiments.

Keywords:
Millepede-IIX-ray crystallographydetector alignmentserial crystallography

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Area of Science:

  • Crystallography
  • High-energy physics
  • Detector physics

Background:

  • Accurate geometrical parameters of X-ray area detectors are crucial for precise serial crystallography.
  • Previous methods for detector calibration suffered from bias and slow convergence.
  • Simultaneous refinement of detector and crystal parameters is necessary to avoid these issues.

Purpose of the Study:

  • To introduce and validate the Millepede algorithm for refining segmented X-ray area detector geometry in serial crystallography.
  • To demonstrate the method's ability to accurately and efficiently correct detector parameters, including tilts and distances.
  • To improve the indexable fraction of data and enable real-time calibration.

Main Methods:

  • Adaptation of the 'Millepede' algorithm, originally from high-energy physics, for serial crystallography.
  • Simultaneous refinement of detector geometrical parameters with individual crystal parameters.
  • Utilizing the structure of least-squares normal equations for rapid computation.

Main Results:

  • Simulated data showed panel shifts within 7% of correct values after one iteration, and near-perfect accuracy after two.
  • A simulated out-of-plane panel rotation was determined to within 0.001°.
  • Experimental data from an X-ray free-electron laser saw the indexable fraction of frames increase from 30% to 91% in one iteration, reaching 96% after further iterations.
  • Geometry updates for 2060 crystals took only 0.819 seconds on desktop hardware.
  • The method was successfully integrated into a real-time feedback system at a synchrotron beamline, correcting a 0.04° detector tilt.

Conclusions:

  • The Millepede method provides a fast, accurate, and practical solution for refining X-ray area detector geometry in serial crystallography.
  • This technique significantly enhances data quality and efficiency in X-ray diffraction experiments.
  • The method's speed and accuracy allow for frequent recalibration, maintaining optimal detector performance without specialized alignment.