Limits...
Fully automatic characterization and data collection from crystals of biological macromolecules.

Svensson O, Malbet-Monaco S, Popov A, Nurizzo D, Bowler MW - Acta Crystallogr. D Biol. Crystallogr. (2015)

Bottom Line: Much of this work is repetitive, and the time spent could be better invested in the interpretation of the results.In order to decrease the need for manual intervention in the most repetitive steps of structural biology projects, initial screening and data collection, a fully automatic system has been developed to mount, locate, centre to the optimal diffraction volume, characterize and, if possible, collect data from multiple cryocooled crystals.Using the capabilities of pixel-array detectors, the system is as fast as a human operator, taking an average of 6 min per sample depending on the sample size and the level of characterization required.

View Article: PubMed Central - HTML - PubMed

Affiliation: European Synchrotron Radiation Facility, 71 Avenue des Martyrs, CS 40220, 38043 Grenoble, France.

ABSTRACT
Considerable effort is dedicated to evaluating macromolecular crystals at synchrotron sources, even for well established and robust systems. Much of this work is repetitive, and the time spent could be better invested in the interpretation of the results. In order to decrease the need for manual intervention in the most repetitive steps of structural biology projects, initial screening and data collection, a fully automatic system has been developed to mount, locate, centre to the optimal diffraction volume, characterize and, if possible, collect data from multiple cryocooled crystals. Using the capabilities of pixel-array detectors, the system is as fast as a human operator, taking an average of 6 min per sample depending on the sample size and the level of characterization required. Using a fast X-ray-based routine, samples are located and centred systematically at the position of highest diffraction signal and important parameters for sample characterization, such as flux, beam size and crystal volume, are automatically taken into account, ensuring the calculation of optimal data-collection strategies. The system is now in operation at the new ESRF beamline MASSIF-1 and has been used by both industrial and academic users for many different sample types, including crystals of less than 20 µm in the smallest dimension. To date, over 8000 samples have been evaluated on MASSIF-1 without any human intervention.

No MeSH data available.


Log-normal distributions of durations for major steps in the automatic treatment of the first 1240 samples processed in 2015 on MASSIF-1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4528805&req=5

fig6: Log-normal distributions of durations for major steps in the automatic treatment of the first 1240 samples processed in 2015 on MASSIF-1.

Mentions: The time taken to process each sample is important as it determines the number of samples that can be processed during scheduled beamtime. This varies depending on the size and shape of the mount and the size of the sample, as these will affect the time taken for the mesh scan and the eventual data-collection time. Fig. 6 ▸ and Table 2 ▸ show the distribution of durations for the major steps in the treatment of all samples processed in the first two months of 2015 (N = 1240) on MASSIF-1. The entire process takes an average of just over 7 min, with the most common time being just over 6 min. The longest steps in the process are the initial mesh scan (average of 101 s) and data collection (averages of 108 and 113 s for EDNA strategies and default collection, respectively). The time taken for default data collection has a very tight distribution, reflecting only small changes in the flux owing to differing ring currents. Data collections based on EDNA strategy calculations are far more varied and often take much less time (most frequently 72 s as opposed to 113 s), but there are also many with much longer data-collection times, showing that longer exposure times can be used when crystal size permits (Table 3 ▸). This will lead to higher signal-to-noise ratios and demonstrates the usefulness of data-collection strategy calculation.


Fully automatic characterization and data collection from crystals of biological macromolecules.

Svensson O, Malbet-Monaco S, Popov A, Nurizzo D, Bowler MW - Acta Crystallogr. D Biol. Crystallogr. (2015)

Log-normal distributions of durations for major steps in the automatic treatment of the first 1240 samples processed in 2015 on MASSIF-1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4528805&req=5

fig6: Log-normal distributions of durations for major steps in the automatic treatment of the first 1240 samples processed in 2015 on MASSIF-1.
Mentions: The time taken to process each sample is important as it determines the number of samples that can be processed during scheduled beamtime. This varies depending on the size and shape of the mount and the size of the sample, as these will affect the time taken for the mesh scan and the eventual data-collection time. Fig. 6 ▸ and Table 2 ▸ show the distribution of durations for the major steps in the treatment of all samples processed in the first two months of 2015 (N = 1240) on MASSIF-1. The entire process takes an average of just over 7 min, with the most common time being just over 6 min. The longest steps in the process are the initial mesh scan (average of 101 s) and data collection (averages of 108 and 113 s for EDNA strategies and default collection, respectively). The time taken for default data collection has a very tight distribution, reflecting only small changes in the flux owing to differing ring currents. Data collections based on EDNA strategy calculations are far more varied and often take much less time (most frequently 72 s as opposed to 113 s), but there are also many with much longer data-collection times, showing that longer exposure times can be used when crystal size permits (Table 3 ▸). This will lead to higher signal-to-noise ratios and demonstrates the usefulness of data-collection strategy calculation.

Bottom Line: Much of this work is repetitive, and the time spent could be better invested in the interpretation of the results.In order to decrease the need for manual intervention in the most repetitive steps of structural biology projects, initial screening and data collection, a fully automatic system has been developed to mount, locate, centre to the optimal diffraction volume, characterize and, if possible, collect data from multiple cryocooled crystals.Using the capabilities of pixel-array detectors, the system is as fast as a human operator, taking an average of 6 min per sample depending on the sample size and the level of characterization required.

View Article: PubMed Central - HTML - PubMed

Affiliation: European Synchrotron Radiation Facility, 71 Avenue des Martyrs, CS 40220, 38043 Grenoble, France.

ABSTRACT
Considerable effort is dedicated to evaluating macromolecular crystals at synchrotron sources, even for well established and robust systems. Much of this work is repetitive, and the time spent could be better invested in the interpretation of the results. In order to decrease the need for manual intervention in the most repetitive steps of structural biology projects, initial screening and data collection, a fully automatic system has been developed to mount, locate, centre to the optimal diffraction volume, characterize and, if possible, collect data from multiple cryocooled crystals. Using the capabilities of pixel-array detectors, the system is as fast as a human operator, taking an average of 6 min per sample depending on the sample size and the level of characterization required. Using a fast X-ray-based routine, samples are located and centred systematically at the position of highest diffraction signal and important parameters for sample characterization, such as flux, beam size and crystal volume, are automatically taken into account, ensuring the calculation of optimal data-collection strategies. The system is now in operation at the new ESRF beamline MASSIF-1 and has been used by both industrial and academic users for many different sample types, including crystals of less than 20 µm in the smallest dimension. To date, over 8000 samples have been evaluated on MASSIF-1 without any human intervention.

No MeSH data available.