title: Room-temperature macromolecular serial crystallography using synchrotron radiation.
abstract: A new approach for collecting data from many hundreds of thousands of microcrystals using X-ray pulses from a free-electron laser has recently been developed. Referred to as serial crystallography, diffraction patterns are recorded at a constant rate as a suspension of protein crystals flows across the path of an X-ray beam. Events that by chance contain single-crystal diffraction patterns are retained, then indexed and merged to form a three-dimensional set of reflection intensities for structure determination. This approach relies upon several innovations: an intense X-ray beam; a fast detector system; a means to rapidly flow a suspension of crystals across the X-ray beam; and the computational infrastructure to process the large volume of data. Originally conceived for radiation-damage-free measurements with ultrafast X-ray pulses, the same methods can be employed with synchrotron radiation. As in powder diffraction, the averaging of thousands of observations per Bragg peak may improve the ratio of signal to noise of low-dose exposures. Here, it is shown that this paradigm can be implemented for room-temperature data collection using synchrotron radiation and exposure times of less than 3 ms. Using lysozyme microcrystals as a model system, over 40 000 single-crystal diffraction patterns were obtained and merged to produce a structural model that could be refined to 2.1 Å resolution. The resulting electron density is in excellent agreement with that obtained using standard X-ray data collection techniques. With further improvements the method is well suited for even shorter exposures at future and upgraded synchrotron radiation facilities that may deliver beams with 1000 times higher brightness than they currently produce.
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany.
authors: Francesco
authors: F
authors: Stellato
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany ; Institute of Biochemistry and Molecular Biology, University of Hamburg , Hamburg 22607, Germany.
authors: Dominik
authors: D
authors: Oberthür
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany.
authors: Mengning
authors: M
authors: Liang
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany.
authors: Richard
authors: R
authors: Bean
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany.
authors: Cornelius
authors: C
authors: Gati
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany.
authors: Oleksandr
authors: O
authors: Yefanov
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany.
authors: Anton
authors: A
authors: Barty
authors: Photon Science, DESY , Hamburg 22607, Germany.
authors: Anja
authors: A
authors: Burkhardt
authors: Photon Science, DESY , Hamburg 22607, Germany.
authors: Pontus
authors: P
authors: Fischer
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany ; Department of Physics, University of Hamburg , Luruper Chaussee 149, Hamburg 22607, Germany.
authors: Lorenzo
authors: L
authors: Galli
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany.
authors: Richard A
authors: RA
authors: Kirian
authors: Photon Science, DESY , Hamburg 22607, Germany.
authors: Jan
authors: J
authors: Meyer
authors: Photon Science, DESY , Hamburg 22607, Germany.
authors: Saravanan
authors: S
authors: Panneerselvam
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany ; European XFEL GmbH, Albert Einstein Ring 19, Hamburg 22761, Germany.
authors: Chun Hong
authors: CH
authors: Yoon
authors: Moscow Institute of Physics and Technology, 141700 Moscow, Russian Federation.
authors: Fedor
authors: F
authors: Chervinskii
authors: Department of Physics, University of York , Heslington, York YO10 5DD, UK.
authors: Emily
authors: E
authors: Speller
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany.
authors: Thomas A
authors: TA
authors: White
authors: Institute of Biochemistry and Molecular Biology, University of Hamburg , Hamburg 22607, Germany ; Center for Ultrafast Imaging, Luruper Chaussee 149, Hamburg 22761, Germany.
authors: Christian
authors: C
authors: Betzel
authors: Photon Science, DESY , Hamburg 22607, Germany.
authors: Alke
authors: A
authors: Meents
authors: Center for Free Electron Laser Science, DESY , Notkestrasse 85, Hamburg 22607, Germany ; Department of Physics, University of Hamburg , Luruper Chaussee 149, Hamburg 22607, Germany ; Center for Ultrafast Imaging, Luruper Chaussee 149, Hamburg 22761, Germany.
authors: Henry N
authors: HN
authors: Chapman
keywords: CrystFEL; microfocus beamline; radiation damage; room-temperature protein crystallography; serial crystallography