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TitleSingle versus dual-axis cryo-electron tomography of microtubules assembled in vitro: limits and perspectives.
Journal, issue, pagesJ Struct Biol, Vol. 181, Issue 2, Page 169-178, Year 2013
Publish dateNov 22, 2012
AuthorsAudrey Guesdon / Sophie Blestel / Charles Kervrann / Denis Chrétien /
PubMed AbstractSingle-axis cryo-electron tomography of vitrified specimens has become a method of choice to reconstruct in three dimensions macromolecular assemblies in their cellular context or prepared from ...Single-axis cryo-electron tomography of vitrified specimens has become a method of choice to reconstruct in three dimensions macromolecular assemblies in their cellular context or prepared from purified components. Here, we asked how a dual-axis acquisition scheme would improve three-dimensional reconstructions of microtubules assembled in vitro. We show that in single-axis tomograms, microtubules oriented close to the perpendicular of the tilt axis display diminished contrast, and ultimately transform into sets of parallel lines oriented in the direction of the electron beam when observed in cross-section. Analysis of their three-dimensional Fourier transform indicates that this imaging artifact is due to a decrease in the angular sampling of their equatorial components. Although the second orthogonal series does not fully complement the first one at the specimen level due to increased radiation damage, it still allows elongated features oriented in any directions to be correctly reconstructed, which might be essential for highly heterogeneous specimens such as cells.
External linksJ Struct Biol / PubMed:23178680
MethodsEM (tomography)
Resolution40.0 Å
Structure data

EMDB-2185:
Single-axis cryo-electron tomogram of microtubules assembled in vitro
Method: EM (tomography) / Resolution: 40.0 Å

EMDB-2186:
Dual-axis cryo-electron tomogram of microtubules assembled in vitro
Method: EM (tomography) / Resolution: 40.0 Å

EMDB-2187:
Dual-axis cryo-electron tomogram of microtubules assembled in vitro
Method: EM (tomography) / Resolution: 40.0 Å

Source
  • Sus scrofa (pig)

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