A crystalline sample of CrLOV1 was optimized for serial crystallography. Time-resolved serial synchrotron crystallography provides high-resolution insights into structural changes of CrLOV1 from Δt = 2.5 ms up to Δt = 95 ms post-photoactivation, resolving the geometry of the thio­ether adduct and alteration of the C-terminal region implicated in the signal transduction.

The HARE chip and a toolbox of helpful instrumentation for time-resolved serial synchrotron crystallography are presented.

This review constitutes an overview of the current status of time-resolved crystallography performed at synchrotrons and XFELs on timescales ranging from femtoseconds to minutes. Methods, potential biases, instruments and examples are presented and compared with those for the cryo-trapping of reaction-intermediate states.

The photo-reaction of the LOV1 domain of the Chlamydomonas reinhardtii phototropin is investigated by room-temperature time-resolved serial crystallography. A covalent adduct forms between the C4a atom of the central flavin-mononucleotide chromophore and a protein cysteine. The structure of the adduct is very similar to that of LOV2 determined 23 years ago from the maidenhair fern Phy3.

SFX diffraction data collection at XFELs is becoming more accessible. To extract the most useful biological information from these non-standard experiments, standards for SFX data analysis and structure validation must be redefined.

This work demonstrates how the precipitating properties of ammonium sulfate can be exploited to drive the transition from vapour-diffusion conditions to large-scale batch micro-crystallization and how the specific ammonium sulfate concentration can further be used to fine-tune microcrystal size and size distribution.

A method is presented to perform time-resolved X-ray crystallography with a 63 ms time resolution using a fast pixel detector and partial oscillation data sets. This minimizes the number of crystals (<100) required for a complete experiment.

Serial crystallography relies upon the growth of microcrystals at high concentration. An LCP crystallization protocol for the Blastochloris viridis photosynthetic reaction centre that utilizes seeding from detergent-grown crystals is reported.

Time-resolved serial femtosecond crystallography experiments can be performed with samples delivered on solid supports. Sample consumption is significantly reduced when compared with the popular crystal-delivery system via high-viscosity extrusion.

This article gives an overview of current trends and challenges in serial crystallography, with a focus on results from time-resolved experiments.