Devitrification reduces beam-induced movement in cryo-EM
- As cryo-EM approaches the physical resolution limits imposed by electron optics and radiation damage, it becomes increasingly urgent to address the issues that impede high-resolution structure determination of biological specimens. One of the persistent problems has been beam-induced movement, which occurs when the specimen is irradiated with high-energy electrons. Beam-induced movement results in image blurring and loss of high-resolution information. It is particularly severe for biological samples in unsupported thin films of vitreous water. By controlled devitrification of conventionally plunge-frozen samples, the suspended film of vitrified water was converted into cubic ice, a polycrystalline, mechanically stable solid. It is shown that compared with vitrified samples, devitrification reduces beam-induced movement in the first 5 e Å−2 of an exposure by a factor of ∼4, substantially enhancing the contribution of the initial, minimally damaged frames to a structure. A 3D apoferritin map reconstructed from the first frames of 20 000 particle images of devitrified samples resolved undamaged side chains. Devitrification of frozen-hydrated specimens helps to overcome beam-induced specimen motion in single-particle cryo-EM, as a further step towards realizing the full potential of cryo-EM for high-resolution structure determination.
Author: | Jan-Philip WieferigORCiDGND, Deryck J. MillsORCiD, Werner KühlbrandtORCiDGND |
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URN: | urn:nbn:de:hebis:30:3-757891 |
DOI: | https://doi.org/10.1107/S2052252520016243 |
ISSN: | 2052-2525 |
Parent Title (English): | IUCrJ |
Publisher: | International Union of Crystallography |
Place of publication: | Chester |
Document Type: | Article |
Language: | English |
Year of Completion: | 2021 |
Year of first Publication: | 2021 |
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
Release Date: | 2024/05/02 |
Tag: | beam-induced movement; cryo-EM; devitrification |
Volume: | 8 |
Issue: | 2 |
Page Number: | 9 |
First Page: | 186 |
Last Page: | 194 |
Institutes: | Biochemie, Chemie und Pharmazie / Biochemie und Chemie |
Angeschlossene und kooperierende Institutionen / MPI für Biophysik | |
Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 53 Physik / 530 Physik |
Sammlungen: | Universitätspublikationen |
Licence (German): | Creative Commons - Namensnennung 4.0 |