Author:
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.
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 µm diameter and selecting the most ordered regions of relatively large crystals using X-ray beams of 5–50 µm 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.
Publisher
International Union of Crystallography (IUCr)
Subject
General Medicine,Structural Biology
Cited by
98 articles.
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