title: Acoustic Injectors for Drop-On-Demand Serial Femtosecond Crystallography.
abstract: X-ray free-electron lasers (XFELs) provide very intense X-ray pulses suitable for macromolecular crystallography. Each X-ray pulse typically lasts for tens of femtoseconds and the interval between pulses is many orders of magnitude longer. Here we describe two novel acoustic injection systems that use focused sound waves to eject picoliter to nanoliter crystal-containing droplets out of microplates and into the X-ray pulse from which diffraction data are collected. The on-demand droplet delivery is synchronized to the XFEL pulse scheme, resulting in X-ray pulses intersecting up to 88% of the droplets. We tested several types of samples in a range of crystallization conditions, wherein the overall crystal hit ratio (e.g., fraction of images with observable diffraction patterns) is a function of the microcrystal slurry concentration. We report crystal structures from lysozyme, thermolysin, and stachydrine demethylase (Stc2). Additional samples were screened to demonstrate that these methods can be applied to rare samples.
authors.affiliation: Photon Sciences Division, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Christian G
authors.initials: CG
authors.lastname: Roessler
authors.affiliation: Biosciences Department, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Rakhi
authors.initials: R
authors.lastname: Agarwal
authors.affiliation: Photon Sciences Division, Brookhaven National Laboratory, Upton, NY 11973-5000, USA. Electronic address: mallaire@lbl.gov.
authors.forename: Marc
authors.initials: M
authors.lastname: Allaire
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: Photon Sciences Division, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Babak
authors.initials: B
authors.lastname: Andi
authors.affiliation: Centro de Biotecnologia Molecular Estrutural, Instituto de Física de São Carlos, Universidade de São Paulo, Caixa Postal 369, São Carlos, CEP: 13560-970, Brazil.
authors.forename: José F R
authors.initials: JFR
authors.lastname: Bachega
authors.affiliation: Institut für Biologie, Humboldt-Universität zu Berlin, D-10099 Berlin, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Bommer
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720-8177, USA.
authors.forename: Aaron S
authors.initials: AS
authors.lastname: Brewster
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Browne
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720-8177, USA.
authors.forename: Ruchira
authors.initials: R
authors.lastname: Chatterjee
authors.affiliation: Department of Chemistry, Boston University, Boston, MA 02215-2521, USA.
authors.forename: Eunsun
authors.initials: E
authors.lastname: Cho
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
authors.affiliation: Photon Sciences Division, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Cowan
authors.affiliation: Labcyte Inc., Sunnyvale, CA 94089, USA.
authors.forename: Sammy
authors.initials: S
authors.lastname: Datwani
authors.affiliation: Burnett School of Biomedical Sciences, University of Central Florida, Orlando, FL 32816-2364, USA.
authors.forename: Victor L
authors.initials: VL
authors.lastname: Davidson
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Jim
authors.initials: J
authors.lastname: Defever
authors.affiliation: Labcyte Inc., Sunnyvale, CA 94089, USA.
authors.forename: Brent
authors.initials: B
authors.lastname: Eaton
authors.affiliation: Labcyte Inc., Sunnyvale, CA 94089, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Ellson
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Yiping
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Labcyte Inc., Sunnyvale, CA 94089, USA.
authors.forename: Lucien P
authors.initials: LP
authors.lastname: Ghislain
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: James M
authors.initials: JM
authors.lastname: Glownia
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720-8177, USA.
authors.forename: Guangye
authors.initials: G
authors.lastname: Han
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720-8177, USA.
authors.forename: Johan
authors.initials: J
authors.lastname: Hattne
authors.affiliation: Max-Volmer-Laboratorium für Biophysikalische Chemie, Technische Universität, D-10623 Berlin, Germany; Institut für Biologie, Humboldt-Universität zu Berlin, D-10099 Berlin, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Hellmich
authors.affiliation: Photon Sciences Division, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Annie
authors.initials: A
authors.lastname: Héroux
authors.affiliation: Institut für Biologie, Humboldt-Universität zu Berlin, D-10099 Berlin, Germany.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Ibrahim
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720-8177, USA; Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Jan
authors.initials: J
authors.lastname: Kern
authors.affiliation: Photon Sciences Division, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Anthony
authors.initials: A
authors.lastname: Kuczewski
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Henrik T
authors.initials: HT
authors.lastname: Lemke
authors.affiliation: Department of Chemistry, Boston University, Boston, MA 02215-2521, USA.
authors.forename: Pinghua
authors.initials: P
authors.lastname: Liu
authors.affiliation: Labcyte Inc., Sunnyvale, CA 94089, USA.
authors.forename: Lars
authors.initials: L
authors.lastname: Majlof
authors.affiliation: Labcyte Inc., Sunnyvale, CA 94089, USA.
authors.forename: William M
authors.initials: WM
authors.lastname: McClintock
authors.affiliation: Photon Sciences Division, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Stuart
authors.initials: S
authors.lastname: Myers
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Silke
authors.initials: S
authors.lastname: Nelsen
authors.affiliation: Labcyte Inc., Sunnyvale, CA 94089, USA.
authors.forename: Joe
authors.initials: J
authors.lastname: Olechno
authors.affiliation: Photon Sciences Division, Brookhaven National Laboratory, Upton, NY 11973-5000, USA; Biosciences Department, Brookhaven National Laboratory, Upton, NY 11973-5000, USA. Electronic address: allen.orville@diamond.ac.uk.
authors.forename: Allen M
authors.initials: AM
authors.lastname: Orville
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720-8177, USA.
authors.forename: Nicholas K
authors.initials: NK
authors.lastname: Sauter
authors.affiliation: Photon Sciences Division, Brookhaven National Laboratory, Upton, NY 11973-5000, USA. Electronic address: soares@bnl.gov.
authors.forename: Alexei S
authors.initials: AS
authors.lastname: Soares
authors.affiliation: Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: S Michael
authors.initials: SM
authors.lastname: Soltis
authors.affiliation: Department of Chemistry, Boston University, Boston, MA 02215-2521, USA.
authors.forename: Heng
authors.initials: H
authors.lastname: Song
authors.affiliation: Labcyte Inc., Sunnyvale, CA 94089, USA.
authors.forename: Richard G
authors.initials: RG
authors.lastname: Stearns
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720-8177, USA.
authors.forename: Rosalie
authors.initials: R
authors.lastname: Tran
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA; Department of Chemistry, Stanford University, Stanford, CA 94305-4401, USA.
authors.forename: Yingssu
authors.initials: Y
authors.lastname: Tsai
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720-8177, USA.
authors.forename: Monarin
authors.initials: M
authors.lastname: Uervirojnangkoorn
authors.affiliation: Department of Biochemistry, Molecular Biology & Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Carrie M
authors.initials: CM
authors.lastname: Wilmot
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720-8177, USA.
authors.forename: Vittal
authors.initials: V
authors.lastname: Yachandra
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720-8177, USA.
authors.forename: Junko
authors.initials: J
authors.lastname: Yano
authors.affiliation: Department of Biochemistry, Molecular Biology & Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Erik T
authors.initials: ET
authors.lastname: Yukl
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
authors.forename: Diling
authors.initials: D
authors.lastname: Zhu
authors.affiliation: Institut für Biologie, Humboldt-Universität zu Berlin, D-10099 Berlin, Germany.
authors.forename: Athina
authors.initials: A
authors.lastname: Zouni
mesh_terms: D000162:Acoustics; D018360:Crystallography, X-Ray / Q000295:instrumentation* / Q000379:methods; D004798:Enzymes / Q000737:chemistry*; D008958:Models, Molecular; D009113:Muramidase / Q000737:chemistry; D011487:Protein Conformation; D013820:Thermolysin / Q000737:chemistry
chemical_list: D004798:Enzymes; D009113:Muramidase; D013820:Thermolysin