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Figure 1
The evolution of macromolecular crystallography, told by size and method. Macromolecular X-ray crystallography (MX) gave rise to the first 3D protein structure, of myoglobin, determined by Kendrew and coworkers in 1957 (Kendrew et al., 1958BB77). Crystals for MX structure determination were required to have sizes greater than 100 µm in all dimensions to expose sufficient crystal lattice volumes to the beam (Smyth & Martin, 2000BB133). Study of 5–10 µm crystals was made possible by the development of advanced instrumentation with dedicated microfocus beamlines such as ID23-2 at ESRF (Flot et al., 2010BB50). The advent of single-shot serial crystallography at XFELs opened up the opportunity to study samples (and/or reactions) at physiological temperatures. The first structure reported from this method was lysozyme (Boutet et al., 2012BB14). While serial methods were adopted by synchrotron sources, crystal sizes are typically larger and range between 10 and 50 µm, as seen for the first structure determined by this method (Gati et al., 2014BB58). Sample preparation using methods adapted from cryoEM include MicroED on one hand, with the first published structure in 2013 (Shi et al., 2013BB129), and X-ray crystallography on the other hand, with the VMXm beamline at the Diamond Light Source (Warren et al., 2024BB147), which are designed for the use of samples down to submicrometre sizes. Despite the plethora of methods, macromolecular crystallization currently remains a bottleneck for structural biologists.

IUCrJ
Volume 12| Part 3| May 2025|
ISSN: 2052-2525
https://doi.org/10.1107/S2052252525001484