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Sci Adv. 2019 May 3;5(5):eaav8801. doi: 10.1126/sciadv.aav8801. eCollection 2019 May.

Electrospray sample injection for single-particle imaging with x-ray lasers.

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Laboratory of Molecular Biophysics, Department of Cell and Molecular Biology, Uppsala University, Husargatan 3 (Box 596), SE-75124 Uppsala, Sweden.
European XFEL GmbH, Holzkoppel 4, 22869 Schenefeld, Germany.
Chemistry Research Laboratory, Department of Chemistry, Oxford University, 12 Mansfield Rd, Oxford OX1 3TA, UK.
Centre for BioImaging Sciences, Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117557, Singapore.
Biomedical and X-ray Physics, Department of Applied Physics, AlbaNova University Center, KTH Royal Institute of Technology, SE-10691 Stockholm, Sweden.
ARC Centre of Advanced Molecular Imaging, Department of Chemistry and Physics, La Trobe University, Melbourne, Victoria 3086, Australia.
Division of Scientific Computing, Department of Information Technology, Science for Life Laboratory, Uppsala University, Lägerhyddsvägen 2 (Box 337), SE-751 05 Uppsala, Sweden.
Department of Physics and Astronomy, Uppsala University, Box 516, SE-75120 Uppsala, Sweden.
Institute of Physics, ELI Beamlines, Academy of Sciences of the Czech Republic, Na Slovance 2, CZ-18221 Prague, Czech Republic.
Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan.
Department of Virology, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima 734-8551, Japan.
Cryo-Electron Microscopy, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, Netherlands.
Department of Physics, National University of Singapore, Singapore, Singapore.
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
Chemical Sciences and Engineering Division, Argonne National Laboratory, Argonne, IL 60439, USA.
Department of Physics and Astronomy, Northwestern University, Evanston, IL 60208, USA.
Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
Institut für Quantenoptik, Leibniz Universität Hannover, Welfengarten 1, 30167 Hannover, Germany.
Institut für Optik und Atomare Physik, Technische Universität Berlin, 10623 Berlin, Germany.
Condensed Matter Physics, Department of Physics, Chalmers University of Technology, Gothenburg, Sweden.
NERSC, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.


The possibility of imaging single proteins constitutes an exciting challenge for x-ray lasers. Despite encouraging results on large particles, imaging small particles has proven to be difficult for two reasons: not quite high enough pulse intensity from currently available x-ray lasers and, as we demonstrate here, contamination of the aerosolized molecules by nonvolatile contaminants in the solution. The amount of contamination on the sample depends on the initial droplet size during aerosolization. Here, we show that, with our electrospray injector, we can decrease the size of aerosol droplets and demonstrate virtually contaminant-free sample delivery of organelles, small virions, and proteins. The results presented here, together with the increased performance of next-generation x-ray lasers, constitute an important stepping stone toward the ultimate goal of protein structure determination from imaging at room temperature and high temporal resolution.

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