title: Fixed target combined with spectral mapping: approaching 100% hit rates for serial crystallography.
abstract: The advent of ultrafast highly brilliant coherent X-ray free-electron laser sources has driven the development of novel structure-determination approaches for proteins, and promises visualization of protein dynamics on sub-picosecond timescales with full atomic resolution. Significant efforts are being applied to the development of sample-delivery systems that allow these unique sources to be most efficiently exploited for high-throughput serial femtosecond crystallography. Here, the next iteration of a fixed-target crystallography chip designed for rapid and reliable delivery of up to 11 259 protein crystals with high spatial precision is presented. An experimental scheme for predetermining the positions of crystals in the chip by means of in situ spectroscopy using a fiducial system for rapid, precise alignment and registration of the crystal positions is presented. This delivers unprecedented performance in serial crystallography experiments at room temperature under atmospheric pressure, giving a raw hit rate approaching 100% with an effective indexing rate of approximately 50%, increasing the efficiency of beam usage and allowing the method to be applied to systems where the number of crystals is limited.
authors.affiliation: Department of Physics and Chemistry, University of Toronto, 60 St George Street, Toronto, ON M5S 1A7, Canada.
authors.forename: Saeed
authors.initials: S
authors.lastname: Oghbaey
authors.affiliation: Department of Physics and Chemistry, University of Toronto, 60 St George Street, Toronto, ON M5S 1A7, Canada.
authors.forename: Antoine
authors.initials: A
authors.lastname: Sarracini
authors.affiliation: Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, England.
authors.forename: Helen M
authors.initials: HM
authors.lastname: Ginn
authors.affiliation: Department of Physics and Chemistry, University of Toronto, 60 St George Street, Toronto, ON M5S 1A7, Canada.
authors.forename: Olivier
authors.initials: O
authors.lastname: Pare-Labrosse
authors.affiliation: Department of Biochemistry, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada.
authors.forename: Anling
authors.initials: A
authors.lastname: Kuo
authors.affiliation: Atomically Resolved Dynamics, Max-Planck-Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, Hamburg, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Marx
authors.affiliation: Atomically Resolved Dynamics, Max-Planck-Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, Hamburg, Germany.
authors.forename: Sascha W
authors.initials: SW
authors.lastname: Epp
authors.affiliation: Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 ODE, England.
authors.forename: Darren A
authors.initials: DA
authors.lastname: Sherrell
authors.affiliation: Department of Biochemistry, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada.
authors.forename: Bryan T
authors.initials: BT
authors.lastname: Eger
authors.affiliation: Atomically Resolved Dynamics, Max-Planck-Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, Hamburg, Germany.
authors.forename: Yinpeng
authors.initials: Y
authors.lastname: Zhong
authors.affiliation: Atomically Resolved Dynamics, Max-Planck-Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, Hamburg, Germany.
authors.forename: Rolf
authors.initials: R
authors.lastname: Loch
authors.affiliation: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.
authors.forename: Valerio
authors.initials: V
authors.lastname: Mariani
authors.affiliation: SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Roberto
authors.initials: R
authors.lastname: Alonso-Mori
authors.affiliation: SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Silke
authors.initials: S
authors.lastname: Nelson
authors.affiliation: SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Henrik T
authors.initials: HT
authors.lastname: Lemke
authors.affiliation: Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 ODE, England.
authors.forename: Robin L
authors.initials: RL
authors.lastname: Owen
authors.affiliation: Hamburg Centre for Ultrafast Imaging, University of Hamburg, Hamburg, Germany.
authors.forename: Arwen R
authors.initials: AR
authors.lastname: Pearson
authors.affiliation: Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, England.
authors.forename: David I
authors.initials: DI
authors.lastname: Stuart
authors.affiliation: Department of Biochemistry, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada.
authors.forename: Oliver P
authors.initials: OP
authors.lastname: Ernst
authors.affiliation: Department of Physics and Chemistry, University of Toronto, 60 St George Street, Toronto, ON M5S 1A7, Canada.
authors.forename: Henrike M
authors.initials: HM
authors.lastname: Mueller-Werkmeister
authors.affiliation: Department of Physics and Chemistry, University of Toronto, 60 St George Street, Toronto, ON M5S 1A7, Canada.
authors.forename: R J Dwayne
authors.initials: RJ
authors.lastname: Miller
mesh_terms: D000818:Animals; D003460:Crystallization / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D018360:Crystallography, X-Ray / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D009211:Myoglobin / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D050661:Sperm Whale; D013696:Temperature; D013997:Time Factors
chemical_list: D009211:Myoglobin; D011506:Proteins
keywords: femtosecond time-resolved crystallography; fixed-target sample delivery; high-throughput serial crystallography; room-temperature crystallography