Advanced serial crystallography utilizing high-energy X-rays enhances diffraction efficiency, data quality and structural resolution under room-temperature conditions.

A correction in the paper by Oberthür et al. [(2025). J. Synchrotron Rad. 32, 474–485] is made.

Facilities for structural biology investigations at DESY and the European XFEL are described.

We present a new error-calibration algorithm for reflection intensities in serial crystallography. We reformulate the mathematical basis of the problem and apply different levels of uncertainty to each observed lattice corresponding to its measurement accuracy. These uncertainties are more consistent with theoretical expectations than the Ev11 error-calibration algorithm previously implemented in cctbx.xfel.merge.

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

An automated, low-volume microbatch-under-oil crystallization approach is described that rapidly maps phase diagram boundaries. This approach rationalizes the production of microcrystal suspensions for serial crystallography by explicitly distinguishing metastable from nucleation zones, thereby replacing empirical trial and error with a quantitative guide to sample optimization.

The influence of the X-ray step size on radiation damage occurring in serial data collection from a metalloprotein using sheet-on-sheet fixed-target chips was investigated at fourth-generation synchrotron and XFEL beamlines.

We demonstrate the application of a nitric oxide releasing photocage system for time-resolved serial crystallography studies of two heme containing proteins using a fixed target sample delivery system. Optimal parameters for successful photocage activation and nitric oxide release are explored.

Developments in macromolecular crystallography now allow the use of microcrystals for structural analysis through advanced beamlines and techniques such as microcrystal electron diffraction and room-temperature crystallography. This review addresses methods of matching microcrystal preparation and sample delivery. The use of microcrystals enhances the possibilities in fields such as time-resolved crystallography.

Fluctuation X-ray scattering measures the correlation of scattered X-rays in diffraction experiments. Here, the Bragg peak intensity from fluctuation X-ray scattering correlations is recovered using an iterative algorithm.

The use of electron beam precession in a (scanning) transmission electron microscope for serial crystallography experiments is thoroughly investigated, showing its potential for better crystal structure determination and refinement in the context of 3D electron diffraction.

We introduce a new signal-separation algorithm for precise background assessment and describe its application to single-crystal image compression and serial crystallography data pre-processing.

Fixed targets (`chips') offer efficient, high-throughput microcrystal delivery for serial crystallography at synchrotrons and X-ray free-electron lasers (XFELs). Within this family, sheet-on-sheet (SOS) chips offer noteworthy advantages in cost, adaptability, universality and ease of crystal loading. We describe our latest generation of SOS devices, which are now in active use at both synchrotrons and XFELs.

Serial crystallography (SX) requires efficient processing of numerous diffraction patterns. TORO Indexer is a high-performance indexing solution that operates across various platforms such as GPUs, CPUs and TPUs, offering high processing speed without compromising indexing quality. Its design ensures easy integration into existing software, making it a useful tool for evolving SX techniques with ever-expanding data volumes.