title: High-density grids for efficient data collection from multiple crystals.
abstract: 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 the Blu-Ice/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.
authors.affiliation: Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Elizabeth L
authors.initials: EL
authors.lastname: Baxter
authors.affiliation: Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Aguila
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Roberto
authors.initials: R
authors.lastname: Alonso-Mori
authors.affiliation: Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
authors.forename: Christopher O
authors.initials: CO
authors.lastname: Barnes
authors.affiliation: Art Robbins Instruments, Sunnyvale, CA 94089, USA.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Bonagura
authors.affiliation: Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Winnie
authors.initials: W
authors.lastname: Brehmer
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Axel T
authors.initials: AT
authors.lastname: Brunger
authors.affiliation: Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
authors.forename: Guillermo
authors.initials: G
authors.lastname: Calero
authors.affiliation: The ARC Centre of Excellence in Advanced Molecular Imaging, Monash University, Melbourne, Victoria 3800, Australia.
authors.forename: Tom T
authors.initials: TT
authors.lastname: Caradoc-Davies
authors.affiliation: Physical Bioscences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
authors.forename: Ruchira
authors.initials: R
authors.lastname: Chatterjee
authors.affiliation: Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, CA 94158, USA.
authors.forename: William F
authors.initials: WF
authors.lastname: Degrado
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, CA 94158, USA.
authors.forename: James S
authors.initials: JS
authors.lastname: Fraser
authors.affiliation: Institut für Biologie, Humboldt-Universität zu Berlin, 10099 Berlin, Germany.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Ibrahim
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Jan
authors.initials: J
authors.lastname: Kern
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Brian K
authors.initials: BK
authors.lastname: Kobilka
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Kruse
authors.affiliation: Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Karl M
authors.initials: KM
authors.lastname: Larsson
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Heinrik T
authors.initials: HT
authors.lastname: Lemke
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Artem Y
authors.initials: AY
authors.lastname: Lyubimov
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Aashish
authors.initials: A
authors.lastname: Manglik
authors.affiliation: Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Scott E
authors.initials: SE
authors.lastname: McPhillips
authors.affiliation: Art Robbins Instruments, Sunnyvale, CA 94089, USA.
authors.forename: Erik
authors.initials: E
authors.lastname: Norgren
authors.affiliation: The ARC Centre of Excellence in Advanced Molecular Imaging, Monash University, Melbourne, Victoria 3800, Australia.
authors.forename: Siew S
authors.initials: SS
authors.lastname: Pang
authors.affiliation: Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: S M
authors.initials: SM
authors.lastname: Soltis
authors.affiliation: Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Jinhu
authors.initials: J
authors.lastname: Song
authors.affiliation: Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, CA 94158, USA.
authors.forename: Jessica
authors.initials: J
authors.lastname: Thomaston
authors.affiliation: Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Yingssu
authors.initials: Y
authors.lastname: Tsai
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305, USA.
authors.forename: William I
authors.initials: WI
authors.lastname: Weis
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, CA 94158, USA.
authors.forename: Rahel A
authors.initials: RA
authors.lastname: Woldeyes
authors.affiliation: Physical Bioscences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
authors.forename: Vittal
authors.initials: V
authors.lastname: Yachandra
authors.affiliation: Physical Bioscences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
authors.forename: Junko
authors.initials: J
authors.lastname: Yano
authors.affiliation: Institut für Biologie, Humboldt-Universität zu Berlin, 10099 Berlin, Germany.
authors.forename: Athina
authors.initials: A
authors.lastname: Zouni
authors.affiliation: Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Aina E
authors.initials: AE
authors.lastname: Cohen
mesh_terms: D000818:Animals; D003460:Crystallization / Q000191:economics / Q000295:instrumentation* / Q000379:methods; D018360:Crystallography, X-Ray / Q000191:economics / Q000295:instrumentation* / Q000379:methods; D003625:Data Collection; D004058:Diffusion; D004867:Equipment Design; D006801:Humans; D013696:Temperature; D014835:Volatilization
keywords: XFELs; automation for sample-exchange robots; high-throughput crystallography; sample delivery; serial crystallography