Fixed-target protein serial microcrystallography with an X-ray free electron laser

Mark S. Hunter, Brent Segelke, Marc Messerschmidt, Garth J. Williams, Nadia Zatsepin, Anton Barty, W. Henry Benner, David B. Carlson, Matthew Coleman, Alexander Graf, Stefan P. Hau-Riege, Tommaso Pardini, M. Marvin Seibert, James Evans, Sébastien Boutet, Matthias Frank

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

We present results from experiments at the Linac Coherent Light Source (LCLS) demonstrating that serial femtosecond crystallography (SFX) can be performed to high resolution (∼2.5 Å) using protein microcrystals deposited on an ultra-thin silicon nitride membrane and embedded in a preservation medium at room temperature. Data can be acquired at a high acquisition rate using X-ray free electron laser sources to overcome radiation damage, while sample consumption is dramatically reduced compared to flowing jet methods. We achieved a peak data acquisition rate of 10 Hz with a hit rate of ∼38%, indicating that a complete data set could be acquired in about one 12-hour LCLS shift using the setup described here, or in even less time using hardware optimized for fixed target SFX. This demonstration opens the door to ultra low sample consumption SFX using the technique of diffraction-before- destruction on proteins that exist in only small quantities and/or do not produce the copious quantities of microcrystals required for flowing jet methods.

Original languageEnglish (US)
Article number6026
JournalScientific Reports
Volume4
DOIs
StatePublished - Aug 12 2014

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Lasers
X-Rays
Electrons
Light
Crystallography
Proteins
Radiation
Temperature
Membranes
silicon nitride
Datasets

ASJC Scopus subject areas

  • General

Cite this

Hunter, M. S., Segelke, B., Messerschmidt, M., Williams, G. J., Zatsepin, N., Barty, A., ... Frank, M. (2014). Fixed-target protein serial microcrystallography with an X-ray free electron laser. Scientific Reports, 4, [6026]. https://doi.org/10.1038/srep06026

Fixed-target protein serial microcrystallography with an X-ray free electron laser. / Hunter, Mark S.; Segelke, Brent; Messerschmidt, Marc; Williams, Garth J.; Zatsepin, Nadia; Barty, Anton; Henry Benner, W.; Carlson, David B.; Coleman, Matthew; Graf, Alexander; Hau-Riege, Stefan P.; Pardini, Tommaso; Marvin Seibert, M.; Evans, James; Boutet, Sébastien; Frank, Matthias.

In: Scientific Reports, Vol. 4, 6026, 12.08.2014.

Research output: Contribution to journalArticle

Hunter, MS, Segelke, B, Messerschmidt, M, Williams, GJ, Zatsepin, N, Barty, A, Henry Benner, W, Carlson, DB, Coleman, M, Graf, A, Hau-Riege, SP, Pardini, T, Marvin Seibert, M, Evans, J, Boutet, S & Frank, M 2014, 'Fixed-target protein serial microcrystallography with an X-ray free electron laser', Scientific Reports, vol. 4, 6026. https://doi.org/10.1038/srep06026
Hunter, Mark S. ; Segelke, Brent ; Messerschmidt, Marc ; Williams, Garth J. ; Zatsepin, Nadia ; Barty, Anton ; Henry Benner, W. ; Carlson, David B. ; Coleman, Matthew ; Graf, Alexander ; Hau-Riege, Stefan P. ; Pardini, Tommaso ; Marvin Seibert, M. ; Evans, James ; Boutet, Sébastien ; Frank, Matthias. / Fixed-target protein serial microcrystallography with an X-ray free electron laser. In: Scientific Reports. 2014 ; Vol. 4.
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