Experimental strategies for imaging bioparticles with femtosecond hard X-ray pulses

Benedikt J. Daurer, Kenta Okamoto, Johan Bielecki, Filipe R.N.C. Maia, Kerstin Mühlig, M. Marvin Seibert, Max F. Hantke, Carl Nettelblad, W. Henry Benner, Martin Svenda, Nicuşor Tîmneanu, Tomas Ekeberg, N. Duane Loh, Alberto Pietrini, Alessandro Zani, Asawari D. Rath, Daniel Westphal, Richard Kirian, Salah Awel, Max O. WiedornGijs Van Der Schot, Gunilla H. Carlsson, Dirk Hasse, Jonas A. Sellberg, Anton Barty, Jakob Andreasson, Sébastien Boutet, Garth Williams, Jason Koglin, Inger Andersson, Janos Hajdu, Daniel S.D. Larsson

Research output: Contribution to journalArticlepeer-review

44 Scopus citations


This study explores the capabilities of the Coherent X-ray Imaging Instrument at the Linac Coherent Light Source to image small biological samples. The weak signal from small samples puts a significant demand on the experiment. Aerosolized Omono River virus particles of ∼40 14;nm in diameter were injected into the submicrometre X-ray focus at a reduced pressure. Diffraction patterns were recorded on two area detectors. The statistical nature of the measurements from many individual particles provided information about the intensity profile of the X-ray beam, phase variations in the wavefront and the size distribution of the injected particles. The results point to a wider than expected size distribution (from ∼35 to ∼30014;nm in diameter). This is likely to be owing to nonvolatile contaminants from larger droplets during aerosolization and droplet evaporation. The results suggest that the concentration of nonvolatile contaminants and the ratio between the volumes of the initial droplet and the sample particles is critical in such studies. The maximum beam intensity in the focus was found to be 1.9 × 1012 photons per μm2 per pulse. The full-width of the focus at half-maximum was estimated to be 500 14;nm (assuming 20% beamline transmission), and this width is larger than expected. Under these conditions, the diffraction signal from a sample-sized particle remained above the average background to a resolution of 4.25 14;nm. The results suggest that reducing the size of the initial droplets during aerosolization is necessary to bring small particles into the scope of detailed structural studies with X-ray lasers.

Original languageEnglish (US)
Pages (from-to)251-262
Number of pages12
StatePublished - May 1 2017


  • OmRV
  • Omono River virus
  • X-ray diffraction
  • diffraction before destruction
  • flash X-ray imaging
  • free-electron laser
  • virus

ASJC Scopus subject areas

  • General Chemistry
  • Biochemistry
  • General Materials Science
  • Condensed Matter Physics


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