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TitleZernike phase contrast cryo-electron microscopy and tomography for structure determination at nanometer and subnanometer resolutions.
Journal, issue, pagesStructure, Vol. 18, Issue 8, Page 903-912, Year 2010
Publish dateAug 11, 2010
AuthorsKazuyoshi Murata / Xiangan Liu / Radostin Danev / Joanita Jakana / Michael F Schmid / Jonathan King / Kuniaki Nagayama / Wah Chiu /
PubMed AbstractZernike phase contrast cryo-electron microscopy (ZPC-cryoEM) is an emerging technique that is capable of producing higher image contrast than conventional cryoEM. By combining this technique with ...Zernike phase contrast cryo-electron microscopy (ZPC-cryoEM) is an emerging technique that is capable of producing higher image contrast than conventional cryoEM. By combining this technique with advanced image processing methods, we achieved subnanometer resolution for two biological specimens: 2D bacteriorhodopsin crystal and epsilon15 bacteriophage. For an asymmetric reconstruction of epsilon15 bacteriophage, ZPC-cryoEM can reduce the required amount of data by a factor of approximately 3, compared with conventional cryoEM. The reconstruction was carried out to 13 A resolution without the need to correct the contrast transfer function. New structural features at the portal vertex of the epsilon15 bacteriophage are revealed in this reconstruction. Using ZPC cryo-electron tomography (ZPC-cryoET), a similar level of data reduction and higher resolution structures of epsilon15 bacteriophage can be obtained relative to conventional cryoET. These results show quantitatively the benefits of ZPC-cryoEM and ZPC-cryoET for structural determinations of macromolecular machines at nanometer and subnanometer resolutions.
External linksStructure / PubMed:20696391 / PubMed Central
MethodsEM (subtomogram averaging) / EM (single particle)
Resolution9.0 - 38.0 Å
Structure data

EMDB-5203:
Subtomogram average of epsilon15 using Zernike phase contrast microscopy
Method: EM (subtomogram averaging) / Resolution: 32.0 Å

EMDB-5204:
Average of 50 Epsilon15 subtomograms using electron cryo-tomography
Method: EM (subtomogram averaging) / Resolution: 38.0 Å

EMDB-5206:
Epsilon15 icosahedral reconstruction using Zernike phase contrast electron microscopy
Method: EM (single particle) / Resolution: 9.0 Å

EMDB-5207:
Epsilon15 asymmetric reconstruction using Zernike phase contrast electron microscopy
Method: EM (single particle) / Resolution: 13.0 Å

EMDB-5208:
Epsilon15 asymmetric reconstruction using Zernike phase contrast electron microscopy (100 particles)
Method: EM (single particle) / Resolution: 26.0 Å

EMDB-5209:
Epsilon15 asymmetric reconstruction using conventional cryo-EM
Method: EM (single particle) / Resolution: 13.0 Å

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