| Title: |
Cryo-EM Sample Preparation with Soft-Landing and Laser Flash Melting |
| Authors: |
SV, Barrass; Esser, Tim K.; NJ, Mowry; Eriksson, L.; Hruby, J; Seeley, Laurence T.; Curtis, Wyatt A.; Baker, Lindsay A.; Drabbels, Marcel; Rauschenbach, Stephan; Lorenz, Ulrich J. |
| Publisher Information: |
bioRxiv |
| Publication Year: |
2026 |
| Collection: |
Ecole Polytechnique Fédérale Lausanne (EPFL): Infoscience |
| Description: |
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 demonstrate that proteins can be returned to their native state by depositing a layer of amorphous ice and briefly flash melting the sample to rehydrate the soft-landed proteins. Melting with a 30 µs laser pulse creates a broad ensemble of partially rehydrated conformations. However, a subset of particles recovers their native configuration. This allows us to propose strategies to fully rehydrate the entire ensemble and brings routine sample preparation by soft-landing mass spectrometry within reach. Finally, the experiments also expand the toolbox of microsecond time-resolved cryo-EM for initiating a broader range of protein dynamics. ; LUXS ; LND |
| Document Type: |
report |
| Language: |
unknown |
| Relation: |
#PLACEHOLDER_PARENT_METADATA_VALUE#; 883387; https://infoscience.epfl.ch/handle/20.500.14299/259599 |
| DOI: |
10.1101/2025.06.05.657968 |
| Availability: |
https://doi.org/10.1101/2025.06.05.657968; https://infoscience.epfl.ch/handle/20.500.14299/259599; https://hdl.handle.net/20.500.14299/259599 |
| Accession Number: |
edsbas.466FFC2B |
| Database: |
BASE |