Resolution extension by image summing in serial femtosecond crystallography of two-dimensional membrane-protein crystals

Cecilia M. Casadei, Ching Ju Tsai, Anton Barty, Mark S. Hunter, Nadia Zatsepin, Celestino Padeste, Guido Capitani, W. Henry Benner, Sébastien Boutet, Stefan P. Hau-Riege, Christopher Kupitz, Marc Messerschmidt, John I. Ogren, Tom Pardini, Kenneth J. Rothschild, Leonardo Sala, Brent Segelke, Garth J. Williams, James E. Evans, Xiao Dan LiMatthew Coleman, Bill Pedrini, Matthias Frank

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


Previous proof-of-concept measurements on single-layer two-dimensional membrane-protein crystals performed at X-ray free-electron lasers (FELs) have demonstrated that the collection of meaningful diffraction patterns, which is not possible at synchrotrons because of radiation-damage issues, is feasible. Here, the results obtained from the analysis of a thousand single-shot, room-temperature X-ray FEL diffraction images from two-dimensional crystals of a bacteriorhodopsin mutant are reported in detail. The high redundancy in the measurements boosts the intensity signal-to-noise ratio, so that the values of the diffracted intensities can be reliably determined down to the detector-edge resolution of 4'Å. The results show that two-dimensional serial crystallography at X-ray FELs is a suitable method to study membrane proteins to near-atomic length scales at ambient temperature. The method presented here can be extended to pump-probe studies of optically triggered structural changes on submillisecond timescales in two-dimensional crystals, which allow functionally relevant large-scale motions that may be quenched in three-dimensional crystals.

Original languageEnglish (US)
Pages (from-to)103-117
Number of pages15
StatePublished - 2018


  • free-electron lasers
  • membrane proteins
  • serial crystallography
  • two-dimensional crystals

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry
  • Materials Science(all)
  • Condensed Matter Physics


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