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. 2023 Nov 1;10(Pt 6):729-737.
doi: 10.1107/S2052252523008618.

Kilohertz serial crystallography with the JUNGFRAU detector at a fourth-generation synchrotron source

Filip Leonarski  1 , Jie Nan  2 , Zdenek Matej  2 , Quentin Bertrand  3 , Antonia Furrer  3 , Ishkhan Gorgisyan  2 , Monika Bjelčić  2 , Michal Kepa  3 , Hannah Glover  3 , Viktoria Hinger  1 , Thomas Eriksson  2 , Aleksander Cehovin  2 , Mikel Eguiraun  2 , Piero Gasparotto  4 , Aldo Mozzanica  1 , Tobias Weinert  3 , Ana Gonzalez  2 , Jörg Standfuss  3 , Meitian Wang  1 , Thomas Ursby  2 , Florian Dworkowski  1
Affiliations

Kilohertz serial crystallography with the JUNGFRAU detector at a fourth-generation synchrotron source

Filip Leonarski et al. IUCrJ. .

Abstract

Serial and time-resolved macromolecular crystallography are on the rise. However, beam time at X-ray free-electron lasers is limited and most third-generation synchrotron-based macromolecular crystallography beamlines do not offer the necessary infrastructure yet. Here, a new setup is demonstrated, based on the JUNGFRAU detector and Jungfraujoch data-acquisition system, that enables collection of kilohertz serial crystallography data at fourth-generation synchrotrons. More importantly, it is shown that this setup is capable of collecting multiple-time-point time-resolved protein dynamics at kilohertz rates, allowing the probing of microsecond to second dynamics at synchrotrons in a fraction of the time needed previously. A high-quality complete X-ray dataset was obtained within 1 min from lysozyme microcrystals, and the dynamics of the light-driven sodium-pump membrane protein KR2 with a time resolution of 1 ms could be demonstrated. To make the setup more accessible for researchers, downstream data handling and analysis will be automated to allow on-the-fly spot finding and indexing, as well as data processing.

Keywords: JUNGFRAU detector; X-ray detectors; data networks; fourth-generation synchrotrons; macromolecular crystallography; protein structure; time-resolved serial crystallography.

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Figures

Figure 1
Figure 1
The setup at the BioMAX beamline. Blue represents the JUNGFRAU 4M prototype detector, yellow is the existing MD3 diffractometer, orange is the transient laser triggering setup and green is the mounted HVE.
Figure 2
Figure 2
Components and connection of the Jungfraujoch data-acquisition system used at MAX IV. The following networks were used for the experiment: (gray/black) a network specifically installed for the experiment, (red) InfiniBand fabric for file-system access, (purple) a fast (40 Gbit s−1) Ethernet network for streaming and (blue) a slow (1 Gbit s−1) control network.
Figure 3
Figure 3
(a) A schematic explanation of the data-collection pattern used to record the millisecond datasets. (b) A difference map of F o from bin #21 (probe, 1 ms after illumination) minus F o of bin #8 (dark). (c) A difference map of 1 ms F o (PDB ID 6tk2) minus dark F o (PDB ID 6tk6) recorded at SwissFEL [data from Skopintsev et al. (2020 ▸)]. All the maps are shown at ±3σ and resolution was cut at 2.38 Å.
See this image and copyright information in PMC

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