Cryo-EM sample preparation with soft-landing and laser flash melting
Abstract The preparation of cryo-EM samples by soft-landing mass spectrometry promises to significantly simplify sample optimization, which has remained an important bottleneck in single-particle cryo-electron microscopy (cryo‑EM). However, only compacted configurations are observed after soft-landing since the proteins are dehydrated in the process. Here, we deposit a layer of amorphous ice onto the sample and briefly flash melt it to rehydrate the soft-landed proteins. Melting with a 30 µs laser pulse creates a broad ensemble of partially rehydrated structures. However, a subset of particles returns to a configuration close to the native state. This allows us to propose strategies to fully rehydrate the entire ensemble and brings routine sample preparation by soft-landing mass spectrometry within reach.
Authors
- Lindsay A. Baker (ORCID: https://orcid.org/0000-0001-9578-4263)
- Stephan Rauschenbach (ORCID: https://orcid.org/0000-0001-9382-1948)
- L. Eriksson (ORCID: https://orcid.org/0009-0000-1149-6118)
- Marcel Drabbels (ORCID: https://orcid.org/0000-0001-8611-2684)
- Mowry NJ (ORCID: https://orcid.org/0000-0001-8635-7075)
- Tim K. Esser (ORCID: https://orcid.org/0000-0002-1174-4131)
- Barrass SV (ORCID: https://orcid.org/0000-0001-7319-6268)
- Ulrich J. Lorenz (ORCID: https://orcid.org/0000-0002-8869-5999)
- J Hruby (ORCID: https://orcid.org/0000-0002-5837-9119)
- Laurence T. Seeley (ORCID: https://orcid.org/0000-0003-2968-3725)
- Curtis WA (ORCID: https://orcid.org/0009-0004-9452-5285)
Institutions
- University of Oxford (GB)
- Science Oxford (GB)
- École Polytechnique Fédérale de Lausanne (CH)
Publication Details
- Journal
- Nature Communications
- Published
- 2026-09-15
- DOI
- https://doi.org/10.1038/s41467-026-77723-6
- Primary Topic
- Advanced Electron Microscopy Techniques and Applications
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Science Foundation
- Wellcome Trust
- UK Research and Innovation
- Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
- Horizon 2020 Framework Programme