Sample-efficient time-resolved X-ray crystallography with serial small wedges
- 1European Synchrotron Radiation Facility
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 [A] 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.
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BibTeX
@misc{bukhdruker2026sampleefficient,
title = {{Sample-efficient time-resolved X-ray crystallography with serial small wedges}},
author = {S. Bukhdruker and R. Astashkin and D. Bratanov and N. Caramello and S. Engilberge and C. Baeken and T. Balandin and A. Popov and A. Royant and C. Mueller-Dieckmann and M. Soler-Lopez and E. Bamberg and V. Gordeliy and I. Melnikov},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.04.756486},
url = {https://www.biorxiv.org/content/10.64898/2026.10.04.756486v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Sample-efficient time-resolved X-ray crystallography with serial small wedges
AU - S. Bukhdruker
AU - R. Astashkin
AU - D. Bratanov
AU - N. Caramello
AU - S. Engilberge
AU - C. Baeken
AU - T. Balandin
AU - A. Popov
AU - A. Royant
AU - C. Mueller-Dieckmann
AU - M. Soler-Lopez
AU - E. Bamberg
AU - V. Gordeliy
AU - I. Melnikov
PY - 2026
DA - 2026/10/04
PB - bioRxiv
DO - 10.64898/2026.10.04.756486
UR - https://www.biorxiv.org/content/10.64898/2026.10.04.756486v1
AB - 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 [A] 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.
ER -