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Diffraction cartography: applying microbeams to macromolecular crystallography sample evaluation and data collection.

Authors
  • Bowler, Matthew W
  • Guijarro, Matias
  • Petitdemange, Sebastien
  • Baker, Isabel
  • Svensson, Olof
  • Burghammer, Manfred
  • Mueller-Dieckmann, Christoph
  • Gordon, Elspeth J
  • Flot, David
  • McSweeney, Sean M
  • Leonard, Gordon A
Type
Published Article
Journal
Acta Crystallographica Section D Biological Crystallography
Publisher
International Union of Crystallography
Publication Date
Aug 01, 2010
Volume
66
Issue
Pt 8
Pages
855–864
Identifiers
DOI: 10.1107/S0907444910019591
PMID: 20693684
Source
Medline
License
Unknown

Abstract

Crystals of biological macromolecules often exhibit considerable inter-crystal and intra-crystal variation in diffraction quality. This requires the evaluation of many samples prior to data collection, a practice that is already widespread in macromolecular crystallography. As structural biologists move towards tackling ever more ambitious projects, new automated methods of sample evaluation will become crucial to the success of many projects, as will the availability of synchrotron-based facilities optimized for high-throughput evaluation of the diffraction characteristics of samples. Here, two examples of the types of advanced sample evaluation that will be required are presented: searching within a sample-containing loop for microcrystals using an X-ray beam of 5 microm diameter and selecting the most ordered regions of relatively large crystals using X-ray beams of 5-50 microm in diameter. A graphical user interface developed to assist with these screening methods is also presented. For the case in which the diffraction quality of a relatively large crystal is probed using a microbeam, the usefulness and implications of mapping diffraction-quality heterogeneity (diffraction cartography) are discussed. The implementation of these techniques in the context of planned upgrades to the ESRF's structural biology beamlines is also presented.

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