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TitleMeasuring the optimal exposure for single particle cryo-EM using a 2.6 Å reconstruction of rotavirus VP6.
Journal, issue, pagesElife, Vol. 4, Page e06980, Year 2015
Publish dateMay 29, 2015
AuthorsTimothy Grant / Nikolaus Grigorieff /
PubMed AbstractBiological specimens suffer radiation damage when imaged in an electron microscope, ultimately limiting the attainable resolution. At a given resolution, an optimal exposure can be defined that ...Biological specimens suffer radiation damage when imaged in an electron microscope, ultimately limiting the attainable resolution. At a given resolution, an optimal exposure can be defined that maximizes the signal-to-noise ratio in the image. Using a 2.6 Å resolution single particle cryo-EM reconstruction of rotavirus VP6, determined from movies recorded with a total exposure of 100 electrons/Å(2), we obtained accurate measurements of optimal exposure values over a wide range of resolutions. At low and intermediate resolutions, our measured values are considerably higher than obtained previously for crystalline specimens, indicating that both images and movies should be collected with higher exposures than are generally used. We demonstrate a method of using our optimal exposure values to filter movie frames, yielding images with improved contrast that lead to higher resolution reconstructions. This 'high-exposure' technique should benefit cryo-EM work on all types of samples, especially those of relatively low-molecular mass.
External linksElife / PubMed:26023829 / PubMed Central
MethodsEM (single particle)
Resolution2.6 - 2.8 Å
Structure data

6272

3j9s

EMDB-6272: Single particle cryo-EM structure of rotavirus VP6 at 2.6 Angstroms resolution
PDB-3j9s: Single particle cryo-EM structure of rotavirus VP6 at 2.6 Angstrom resolution
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-6464:
Reconstruction of the T20S proteasome at 2.8 Angstrom resolution using optimal exposure filtering
Method: EM (single particle) / Resolution: 2.8 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-CL:
Unknown entry

ChemComp-CA:
Unknown entry

ChemComp-HOH:
WATER

Source
  • bovine rotavirus strain uk/g6
  • Thermoplasma acidophilum (acidophilic)
KeywordsVIRAL PROTEIN / rotavirus / virus

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