title: Fixed-target protein serial microcrystallography with an x-ray free electron laser.
abstract: We present results from experiments at the Linac Coherent Light Source (LCLS) demonstrating that serial femtosecond crystallography (SFX) can be performed to high resolution (~2.5 Å) using protein microcrystals deposited on an ultra-thin silicon nitride membrane and embedded in a preservation medium at room temperature. Data can be acquired at a high acquisition rate using x-ray free electron laser sources to overcome radiation damage, while sample consumption is dramatically reduced compared to flowing jet methods. We achieved a peak data acquisition rate of 10 Hz with a hit rate of ~38%, indicating that a complete data set could be acquired in about one 12-hour LCLS shift using the setup described here, or in even less time using hardware optimized for fixed target SFX. This demonstration opens the door to ultra low sample consumption SFX using the technique of diffraction-before-destruction on proteins that exist in only small quantities and/or do not produce the copious quantities of microcrystals required for flowing jet methods.
authors.affiliation: Physics Division, Lawrence Livermore National Laboratory, 7000 East Avenue, Mail Stop L-211, Livermore, CA 94550, USA.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Hunter
authors.affiliation: Biosciences and Biotechnology Division, Lawrence Livermore National Laboratory, 7000 East Avenue, Mail Stop L-452, Livermore, CA 94550, USA.
authors.forename: Brent
authors.initials: B
authors.lastname: Segelke
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
authors.forename: Marc
authors.initials: M
authors.lastname: Messerschmidt
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
authors.forename: Garth J
authors.initials: GJ
authors.lastname: Williams
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Nadia A
authors.initials: NA
authors.lastname: Zatsepin
authors.affiliation: Centre for Free-Electron Laser Science, DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Barty
authors.affiliation: Biosciences and Biotechnology Division, Lawrence Livermore National Laboratory, 7000 East Avenue, Mail Stop L-452, Livermore, CA 94550, USA.
authors.forename: W Henry
authors.initials: WH
authors.lastname: Benner
authors.affiliation: Dept. of Molecular and Cellular Biology, University of California, Davis, 1 Shields Ave, Davis, CA, 95616, USA.
authors.forename: David B
authors.initials: DB
authors.lastname: Carlson
authors.affiliation: 1] Biosciences and Biotechnology Division, Lawrence Livermore National Laboratory, 7000 East Avenue, Mail Stop L-452, Livermore, CA 94550, USA [2] Department of Radiation Oncology, University of California at Davis Medical Center, Sacramento, CA 95817.
authors.forename: Matthew
authors.initials: M
authors.lastname: Coleman
authors.affiliation: Physics Division, Lawrence Livermore National Laboratory, 7000 East Avenue, Mail Stop L-211, Livermore, CA 94550, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Graf
authors.affiliation: Physics Division, Lawrence Livermore National Laboratory, 7000 East Avenue, Mail Stop L-211, Livermore, CA 94550, USA.
authors.forename: Stefan P
authors.initials: SP
authors.lastname: Hau-Riege
authors.affiliation: Physics Division, Lawrence Livermore National Laboratory, 7000 East Avenue, Mail Stop L-211, Livermore, CA 94550, USA.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Pardini
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
authors.forename: M Marvin
authors.initials: MM
authors.lastname: Seibert
authors.affiliation: 1] Dept. of Molecular and Cellular Biology, University of California, Davis, 1 Shields Ave, Davis, CA, 95616, USA [2] Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, 3335 Innovation Blvd., Richland, WA, 99354, USA.
authors.forename: James
authors.initials: J
authors.lastname: Evans
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Boutet
authors.affiliation: Physics Division, Lawrence Livermore National Laboratory, 7000 East Avenue, Mail Stop L-211, Livermore, CA 94550, USA.
authors.forename: Matthias
authors.initials: M
authors.lastname: Frank
mesh_terms: D003461:Crystallography / Q000295:instrumentation / Q000379:methods*; D004583:Electrons; D007834:Lasers; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins
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