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TitleCapabilities of the Falcon III detector for single-particle structure determination.
Journal, issue, pagesUltramicroscopy, Vol. 203, Page 145-154, Year 2019
Publish dateJan 28, 2019
AuthorsBoyuan Song / Julian Lenhart / Vanessa Judith Flegler / Cihan Makbul / Tim Rasmussen / Bettina Böttcher /
PubMed AbstractDirect electron detectors are an essential asset for the resolution revolution in electron cryo microscopy of biological objects. The direct detectors provide two modes of data acquisition; the ...Direct electron detectors are an essential asset for the resolution revolution in electron cryo microscopy of biological objects. The direct detectors provide two modes of data acquisition; the counting mode in which single electrons are counted, and the integrating mode in which the signal that arises from the incident electrons is integrated. While counting mode leads to far higher detective quantum efficiency at all spatial frequencies, the integrating mode enables faster data acquisition at higher exposure rates. For optimal throughput at best possible resolution it is important to understand when the better performance in counting mode becomes essential for solving a structure and when the lower detective quantum efficiency in integrating mode can be compensated by increasing the number of particles in the data set. Here, we provide a case study of the Falcon III camera, which has counting mode capability at exposure rates of <0.9 e/Px² and integrating mode capability at exposure rates above 10 e/Px². We found that counting mode gives better resolution for medium sized complexes such as the β-galactosidase (465 kDa) (2.2 Å, 97% of Nyquist vs. 2.4 Å, 89% of Nyquist) with data sets of similar size. However, for larger particles such as Hepatitis B virus capsid like particles (4.8 MDa) we did not find any resolution gain in counting mode.
External linksUltramicroscopy / PubMed:30738626
MethodsEM (helical sym.) / EM (single particle)
Resolution2.2 - 2.4 Å
Structure data

4413

6i5a

EMDB-4413, PDB-6i5a:
Tobacco Mosaic Virus
Method: EM (helical sym.) / Resolution: 2.3 Å

EMDB-4414:
beta-galactosidase (integrating mode)
Method: EM (single particle) / Resolution: 2.4 Å

EMDB-4415:
beta-galactosidase (counting mode)
Method: EM (single particle) / Resolution: 2.2 Å

EMDB-4416:
F97L counting mode
Method: EM (single particle) / Resolution: 2.36 Å

EMDB-4417:
F97L integrating mode
Method: EM (single particle) / Resolution: 2.38 Å

Source
  • TMV (virus)
  • Escherichia coli (E. coli)
  • Hepatitis B virus
  • tobacco mosaic virus (strain vulgare)
KeywordsVIRUS / TMV / helical virus / coat protein / plant pathogen

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