Double-flow focused liquid injector for efficient serial femtosecond crystallography

Sci Rep. 2017 Mar 16;7:44628. doi: 10.1038/srep44628.

Abstract

Serial femtosecond crystallography requires reliable and efficient delivery of fresh crystals across the beam of an X-ray free-electron laser over the course of an experiment. We introduce a double-flow focusing nozzle to meet this challenge, with significantly reduced sample consumption, while improving jet stability over previous generations of nozzles. We demonstrate its use to determine the first room-temperature structure of RNA polymerase II at high resolution, revealing new structural details. Moreover, the double-flow focusing nozzles were successfully tested with three other protein samples and the first room temperature structure of an extradiol ring-cleaving dioxygenase was solved by utilizing the improved operation and characteristics of these devices [corrected].

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Computer Simulation
  • Crystallography / instrumentation*
  • RNA Polymerase II / chemistry
  • Rheology / instrumentation*
  • Saccharomyces cerevisiae / enzymology
  • Temperature
  • Time Factors
  • X-Ray Diffraction

Substances

  • RNA Polymerase II