期刊
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY
卷 72, 期 -, 页码 2-11出版社
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S2059798315020847
关键词
XFELs; high-throughput crystallography; serial crystallography; sample delivery; automation for sample-exchange robots
资金
- US Department of Energy, Office of Basic Energy Sciences [DE-AC02-76SF00515]
- LCLS Ultrafast Science Instruments project - US Department of Energy, Office of Basic Energy Sciences
- US Department of Energy, Office of Science, Office of Basic Energy Sciences [DE-AC02-76SF00515]
- US Department of Energy Office of Biological and Environmental Research
- National Institutes of Health (NIH), National Institute of General Medical Sciences [P41GM103393]
- NSF Graduate Research Fellowship
- NIH [GM110580]
- NSF [STC-1231306]
- Office of Science, OBES, Chemical Sciences, Geosciences and Biosciences CSGB of the DOE [DE-AC02-05CH11231]
- National Institutes of Health (NIH) [GM055302, GM110501]
- DFG-Cluster of Excellence 'UniCat' coordinated by Technische Universitat at Berlin [Sfb1078, TPA5]
- Human Frontiers Science Project [RGP0063/2013 310]
Higher throughput methods to mount and collect data from multiple small and radiation-sensitive crystals are important to support challenging structural investigations using microfocus synchrotron beamlines. Furthermore, efficient sample-delivery methods are essential to carry out productive femtosecond crystallography experiments at X-ray free-electron laser (XFEL) sources such as the Linac Coherent Light Source (LCLS). To address these needs, a high-density sample grid useful as a scaffold for both crystal growth and diffraction data collection has been developed and utilized for efficient goniometer-based sample delivery at synchrotron and XFEL sources. A single grid contains 75 mounting ports and fits inside an SSRL cassette or uni-puck storage container. The use of grids with an SSRL cassette expands the cassette capacity up to 7200 samples. Grids may also be covered with a polymer film or sleeve for efficient room-temperature data collection from multiple samples. New automated routines have been incorporated into theBlu-Ice Blu-Ice/DCSS DCSS experimental control system to support grids, including semi-automated grid alignment, fully automated positioning of grid ports, rastering and automated data collection. Specialized tools have been developed to support crystallization experiments on grids, including a universal adaptor, which allows grids to be filled by commercial liquid-handling robots, as well as incubation chambers, which support vapor-diffusion and lipidic cubic phase crystallization experiments. Experiments in which crystals were loaded into grids or grown on grids using liquid-handling robots and incubation chambers are described. Crystals were screened at LCLS-XPP and SSRL BL12-2 at room temperature and cryogenic temperatures.
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