Foam film vitrification for cryo-EM

Yue Zhang (), Biplob Nandy, Kasim Sader, Christopher J. Russo () and Jan Löwe ()
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Yue Zhang: Medical Research Council (MRC) Laboratory of Molecular Biology
Biplob Nandy: Medical Research Council (MRC) Laboratory of Molecular Biology
Kasim Sader: AstraZeneca
Christopher J. Russo: Medical Research Council (MRC) Laboratory of Molecular Biology
Jan Löwe: Medical Research Council (MRC) Laboratory of Molecular Biology

Nature Communications, 2025, vol. 16, issue 1, 1-12

Abstract: Abstract Electron cryomicroscopy (cryo-EM) has revolutionised structural biology, enhancing applicability, size limits and speed. Despite these successes, cryo-EM sample preparation remains a major bottleneck for routinely achieving high-resolution structures through single particle analysis. Challenges such as inconsistent ice thicknesses, air-water interface interactions and preferred particle orientation persist. Here, we introduce a blot-free vitrification method that uses free-standing surfactant-stabilised foam films to address some of these issues. The method achieves uniform ice thicknesses, enables thickness control of the foam film prior to vitrification, and for some specimens enhances orientation distribution efficiency. Furthermore, it reduces particle adsorption to carbon foil on the specimen support. The method simplifies cryo-EM specimen preparation, offering improved control over ice thickness and particle orientation, to help streamline and accelerate structure determination.

Date: 2025
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DOI: 10.1038/s41467-025-61270-7

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