Dose, exposure time and resolution in serial X-ray crystallography

D. Starodub, Peter Rez, G. Hembree, M. Howells, D. Shapiro, H. N. Chapman, Petra Fromme, Kevin Schmidt, Uwe Weierstall, R. B. Doak, John Spence

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The resolution of X-ray diffraction microscopy is limited by the maximum dose that can be delivered prior to sample damage. In the proposed serial crystallography method, the damage problem is addressed by distributing the total dose over many identical hydrated macromolecules running continuously in a single-file train across a continuous X-ray beam, and resolution is then limited only by the available molecular and X-ray fluxes and molecular alignment. Orientation of the diffracting molecules is achieved by laser alignment. The incident X-ray fluence (energy/area) is evaluated that is required to obtain a given resolution from (i) an analytical model, giving the count rate at the maximum scattering angle for a model protein, (ii) explicit simulation of diffraction patterns for a GroEL-GroES protein complex, and (iii) the spatial frequency cut-off of the transfer function following iterative solution of the phase problem, and reconstruction of an electron density map in the projection approximation. These calculations include counting shot noise and multiple starts of the phasing algorithm. The results indicate counting time and the number of proteins needed within the beam at any instant for a given resolution and X-ray flux. An inverse fourth-power dependence of exposure time on resolution is confirmed, with important implications for all coherent X-ray imaging. It is found that multiple single-file protein beams will be needed for sub-nanometer resolution on current third-generation synchrotrons, but not on fourth-generation designs, where reconstruction of secondary protein structure at a resolution of 7 Å should be possible with relatively short exposures.

Original languageEnglish (US)
Pages (from-to)62-73
Number of pages12
JournalJournal of Synchrotron Radiation
Volume15
Issue number1
DOIs
StatePublished - Dec 18 2007

Fingerprint

X ray crystallography
crystallography
Proteins
X rays
dosage
proteins
x rays
files
Fluxes
Shot noise
Crystallography
counting
Molecular orientation
alignment
Synchrotrons
Macromolecules
damage
Crystal orientation
Diffraction patterns
Carrier concentration

Keywords

  • Coherent scattering
  • Phase retrieval
  • Protein structure
  • Transfer function

ASJC Scopus subject areas

  • Instrumentation
  • Atomic and Molecular Physics, and Optics
  • Radiation
  • Physics and Astronomy (miscellaneous)

Cite this

Dose, exposure time and resolution in serial X-ray crystallography. / Starodub, D.; Rez, Peter; Hembree, G.; Howells, M.; Shapiro, D.; Chapman, H. N.; Fromme, Petra; Schmidt, Kevin; Weierstall, Uwe; Doak, R. B.; Spence, John.

In: Journal of Synchrotron Radiation, Vol. 15, No. 1, 18.12.2007, p. 62-73.

Research output: Contribution to journalArticle

Starodub, D. ; Rez, Peter ; Hembree, G. ; Howells, M. ; Shapiro, D. ; Chapman, H. N. ; Fromme, Petra ; Schmidt, Kevin ; Weierstall, Uwe ; Doak, R. B. ; Spence, John. / Dose, exposure time and resolution in serial X-ray crystallography. In: Journal of Synchrotron Radiation. 2007 ; Vol. 15, No. 1. pp. 62-73.
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