Small but mighty: the power of microcrystals in structural biology

DOI: 10.1107/S2052252525001484

Authors: Courtney J. Tremlett (University of Exeter) , Jack Stubbs (University of Southampton; Diamond Light Source) , William S. Stuart (University of Exeter; Defence Science and Technology Laboratoy) , Patrick D. Shaw Stewart (Douglas Instruments Ltd) , Jonathan West (University of Southampton) , Allen M. Orville (Diamond Light Source; Research Complex at Harwell) , Ivo Tews (University of Southampton) , Nicholas J. Harmer (University of Exeter)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Iucrj , VOL 12

State: Published (Approved)
Published: May 2025

Abstract: Advancements in macromolecular crystallography, driven by improved sources and cryocooling techniques, have enabled the use of increasingly smaller crystals for structure determination, with microfocus beamlines now widely accessible. Initially developed for challenging samples, these techniques have culminated in advanced beamlines such as VMXm. Here, an in vacuo sample environment improves the signal-to-noise ratio in X-ray diffraction experiments, and thus enables the use of submicrometre crystals. The advancement of techniques such as microcrystal electron diffraction (MicroED) for atomic-level insights into charged states and hydrogen positions, along with room-temperature crystallography to observe physiological states via serial crystallography, has driven a resurgence in the use of microcrystals. Reproducibly preparing small crystals, especially from samples that typically yield larger crystals, requires considerable effort, as no one singular approach guarantees optimal crystals for every technique. This review discusses methods for generating such small crystals, including mechanical crushing and batch crystallization with seeding, and evaluates their compatibility with microcrystal data-collection modalities. Additionally, we examine sample-delivery methods, which are crucial for selecting appropriate crystallization strategies. Establishing reliable protocols for sample preparation and delivery opens new avenues for macromolecular crystallography, particularly in the rapidly progressing field of time-resolved crystallography.

Journal Keywords: microcrystals; MicroED; serial crystallography; time-resolved studies; seeding; phase diagrams; sample preparation; sample delivery

Subject Areas: Biology and Bio-materials, Technique Development


Technical Areas:

Added On: 16/03/2025 08:57

Discipline Tags:

Technique Development - Life Sciences & Biotech Structural biology Life Sciences & Biotech

Technical Tags: