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TitleCryo-EM single particle analysis with the Volta phase plate.
Journal, issue, pagesElife, Vol. 5, Year 2016
Publish dateMar 7, 2016
AuthorsRadostin Danev / Wolfgang Baumeister /
PubMed AbstractWe present a method for in-focus data acquisition with a phase plate that enables near-atomic resolution single particle reconstructions. Accurate focusing is the determining factor for obtaining ...We present a method for in-focus data acquisition with a phase plate that enables near-atomic resolution single particle reconstructions. Accurate focusing is the determining factor for obtaining high quality data. A double-area focusing strategy was implemented in order to achieve the required precision. With this approach we obtained a 3.2 Å resolution reconstruction of the Thermoplasma acidophilum 20S proteasome. The phase plate matches or slightly exceeds the performance of the conventional defocus approach. Spherical aberration becomes a limiting factor for achieving resolutions below 3 Å with in-focus phase plate images. The phase plate could enable single particle analysis of challenging samples in terms of small size, heterogeneity and flexibility that are difficult to solve by the conventional defocus approach.
External linksElife / PubMed:26949259 / PubMed Central
MethodsEM (single particle)
Resolution3.1 - 3.2 Å
Structure data

EMDB-3347:
Volta phase plate 3.2A resolution reconstruction of the T20S proteasome
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-3348:
cryo-EM structure of T20S proteasome
Method: EM (single particle) / Resolution: 3.1 Å

Source
  • Thermoplasma acidophilum (acidophilic)

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