Year: 2026
Type: preprint
Abstract: Time-resolved crystallography can be inaccessible for membrane proteins and other difficult-to-crystallize targets for which suitable crystals are scarce. Here we present an acquisition strategy that records consecutive diffraction images during a small crystal rotation synchronized to a single activation pulse. Matching image segments are merged across crystals to reconstruct successive time windows, enabling a single crystal ensemble to capture an extended structural trajectory. Using the Mesh&Collect workflow at a synchrotron, we obtained ten consecutive 7.5-ms time windows at 2 Å resolution from 1,255 microcrystals of a bacteriorhodopsin triple mutant at room temperature. An additional 790 crystals extended the observation window to 500 ms. Rotation-based processing, retrospective time binning and dose control maximize information extracted from each crystal, while compatible measurements can be pooled across acquisitions. This approach makes millisecond time-resolved crystallography practical for crystal-limited targets at synchrotrons, with further development potentially extending the temporal resolution down to 1 ms.
Institutions: Centre National de la Recherche Scientifique, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, European Synchrotron Radiation Facility, CEA Grenoble, Institut de Biologie Structurale, Eindhoven University of Technology, Forschungszentrum Jülich, National Taiwan University, Max Planck Institute of Biophysics
Language: English
Subfield: Materials Chemistry
Field: Materials Science
Domain: Physical Sciences
Open Access status: green
OpenAlex ID: W7221119977