Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers
Susannah Holmes, Henry Kirkwood, Richard Bean, Klaus Giewekemeyer, Andrew V. Martin, Marjan Hadian-Jazi, Max O. Wiedorn, Dominik Oberthür, Hugh Marman, Luigi Adriano, Nasser Al-Qudami, S. Bajt, Imrich Barák, Sadia Bari, Johan Bielecki, Sándor Brockhauser, Matthew A. Coleman, Francisco Cruz-Mazo, Cyril Danilevski, Katerina Dörner, Alfonso M. Gañán‐Calvo, Rita Graceffa, Hans Fanghor, Michaël Heymann, Matthias Frank, Alexander Kaukher, Yoonhee Kim, Boštjan Kobe, J. Knoška, Torsten Laurus, Romain Letrun, Luis Maia, M. Messerschmidt, Markus Metz, Thomas Michelat, Grant Mills, С. Л. Молодцов, Diana C. F. Monteiro, Andrew J. Morgan, Astrid Münnich, Gisel E. Peña Murillo, Gianpietro Previtali, Adam Round, Tokushi Sato, Robin Schubert, Joachim Schulz, Megan L. Shelby, Carolin Seuring, Jonas A. Sellberg, Marcin Sikorski, A. Silenzi, Stephan Stern, Jola Sztuk-Dambietz, J. Szuba, Martin Trebbin, Patrick Vagovic, Thomas Ve, Britta Weinhausen, K. Wrona, P. Lourdu Xavier, Xu Chen, Oleksandr Yefanov, K. Nugent, Henry N. Chapman, Adrian P. Mancuso⋈, Anton Barty, Brian Abbey, Connie Darmanin
Abstract
The European X-ray Free Electron Laser (XFEL) and Linac Coherent Light Source (LCLS) II are extremely intense sources of X-rays capable of generating Serial Femtosecond Crystallography (SFX) data at megahertz (MHz) repetition rates. Previous work has shown that it is possible to use consecutive X-ray pulses to collect diffraction patterns from individual crystals. Here, we exploit the MHz pulse structure of the European XFEL to obtain two complete datasets from the same lysozyme crystal, first hit and the second hit, before it exits the beam. The two datasets, separated by <1 µs, yield up to 2.1 Å resolution structures. Comparisons between the two structures reveal no indications of radiation damage or significant changes within the active site, consistent with the calculated dose estimates. This demonstrates MHz SFX can be used as a tool for tracking sub-microsecond structural changes in individual single crystals, a technique we refer to as multi-hit SFX.