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TitleAdvances in Single-Particle Electron Cryomicroscopy Structure Determination applied to Sub-tomogram Averaging.
Journal, issue, pagesStructure, Vol. 23, Issue 9, Page 1743-1753, Year 2015
Publish dateSep 1, 2015
AuthorsTanmay A M Bharat / Christopher J Russo / Jan Löwe / Lori A Passmore / Sjors H W Scheres /
PubMed AbstractRecent innovations in specimen preparation, data collection, and image processing have led to improved structure determination using single-particle electron cryomicroscopy (cryo-EM). Here we explore ...Recent innovations in specimen preparation, data collection, and image processing have led to improved structure determination using single-particle electron cryomicroscopy (cryo-EM). Here we explore some of these advances to improve structures determined using electron cryotomography (cryo-ET) and sub-tomogram averaging. We implement a new three-dimensional model for the contrast transfer function, and use this in a regularized likelihood optimization algorithm as implemented in the RELION program. Using direct electron detector data, we apply both single-particle analysis and sub-tomogram averaging to analyze radiation-induced movements of the specimen. As in single-particle cryo-EM, we find that significant sample movements occur during tomographic data acquisition, and that these movements are substantially reduced through the use of ultrastable gold substrates. We obtain a sub-nanometer resolution structure of the hepatitis B capsid, and show that reducing radiation-induced specimen movement may be central to attempts at further improving tomogram quality and resolution.
External linksStructure / PubMed:26256537 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution8.1 Å
Structure data

EMDB-3015:
Using recent advances in single-particle electron cryomicroscopy structure determination for sub-tomogram averaging
Method: EM (subtomogram averaging) / Resolution: 8.1 Å

Source
  • Hepatitis B virus

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