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Single versus dual-axis cryo-electron tomography of microtubules assembled in vitro: limits and perspectives.

Abstract：

: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.

journal_name

J Struct Biol

journal_title

Journal of structural biology

authors

Guesdon A,Blestel S,Kervrann C,Chrétien D

doi

10.1016/j.jsb.2012.11.004

subject

Has Abstract

pub_date

2013-02-01 00:00:00

pages

169-78

issue

2

eissn

1047-8477

issn

1095-8657

pii

S1047-8477(12)00316-4

journal_volume

181

pub_type

杂志文章

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