MASSIF-1: fully automatic data collection
The ESRF and EMBL-Grenoble have long worked together on the automation of experiments in macromolecular crystallography (MX). This has now reached its peak in the form of a new beamline on MASSIF-1 (ID30A-1) at the ESRF that fully automates the process of mounting, locating, centring to the optimal diffraction volume, characterising and collecting data, if possible, from multiple cryo-cooled crystals. The EMBL-Grenoble beamline scientist for MASSIF-1 is Matthew Bowler, a member of the Synchrotron Crystallography Team, who commissioned the beamline and has been involved in the MASSIF project since its inception.
MASSIF-1 is a unique facility for the high throughput, fully automatic characterisation and data collection of macromolecular crystals. The new service is not designed to replace user visits to the synchrotron, but rather to do the hard work of screening crystals or collecting routine data sets through the night, freeing researchers to spend more time on challenging data collection problems and study of the underlying biology. These services are hugely popular, with hundreds of samples processed weekly, ranging from initial hits from crystallisation experiments to large-scale data set collection for drug discovery programmes. The automatic routines developed are often able to locate crystals more effectively than the human eye and in many cases have obtained higher resolution data sets, as all positions within a sample can be evaluated for diffraction quality.
Beam time is booked flexibly and samples then enter a queuing system. Users interact with the beamline by describing experimental requirements that are used by the beamline software to set data collection parameters, via ISPyB, where results can also be viewed and downloaded. These services are made possible by RoboDiff – a new ESRF-developed sample changer that also acts as a goniometer, a highly intense X-ray beam (3 x 1012 ph/sec in 100 x 50 µm2) and complex workflows that fully evaluate samples, centre the best volumes and collect diffraction data sets optimised for maximum resolution with minimised radiation damage. As MASSIF-1 is fully automatic, data are collected for the first time in a consistent manner and should allow the accumulation and comparison of a large amount of information that was previously unknown, including the exact dimensions of crystals and deeper information about their quality. Once the beamline has been running for an extended period, it will provide a treasure trove of additional information to feed back into crystallisation experiments and the software used to collect the data.
MASSIF-1 is run as a collaboration between the EMBL-Grenoble Synchrotron Crystallography Team and the ESRF Structural Biology Group. See the MASSIF1 webpage for more information on the technical details and the ESRF website for application details.
Bowler MW, Svensson O & Nurizzo D. (2016): Fully automatic macromolecular crystallography: the impact of MASSIF-1 on the optimum acquisition and quality of data, Crystallography Reviews, doi:10.1080/0889311X.2016.1155050
Nurizzo D, Bowler MW et al. (2016) RoboDiff: combining a sample changer and goniometer for highly automated macromolecular crystallography experiments, Acta Cryst D72, 966-975, doi:10.1107/S205979831601158X
Svensson O, Monaco S, Popov AN, Nurizzo D, Bowler MW (2015). The fully automatic characterization and data collection from crystals of biological macromolecules, Acta Cryst. D71, 1757-1767, doi:10.1107/S1399004715011918
Bowler MW, Nurizzo D et al. (2015). MASSIF-1: A beamline dedicated to the fully automatic characterisation and data collection from crystals of biological macromolecules, J. Synchrotron Rad. 22, 1540-1547, doi:10.1107/S1600577515016604
Thierry E, Guilligay D, Kosinski J, Bock T, Gaudon S, Round A, Pflug A, Hengrung N, El Omari K, Baudin F, Hart DJ, Beck M, Cusack S (2016). Influenza Polymerase Can Adopt an Alternative Configuration Involving a Radical Repacking of PB2 Domains, Molecular Cell. 61:125-37, doi:10.1016/j.molcel.2015.11.016
Neudegger T, Verghese J, Hayer-Hart M, Hartl FU, Bracher A (2016). Structure of human heat-shock transcription factor 1 in complex with DNA, Nat Struct Mol Biol, doi:10.1038/nsmb.3149
Kharde S, Calviño FR, Gumiero A, Wild K, Sinning I (2015). The structure of Rpf2-Rrs1 explains its role in ribosome biogenesis, Nucleic Acids Res. 43, 7083–95, doi:10.1093/nar/gkv640