A "flip-flop" rotation stage for routine dual-axis electron cryotomography

Cristina V. Iancu, Elizabeth R. Wright, Jordan Benjamin, William F. Tivol, D. Prabha Dias, Gavin E. Murphy, Robert C. Morrison, J. Bernard Heymann, Grant J. Jensen

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Abstract

Electron cryotomography can be used to solve the three-dimensional structures of individual large macromolecules, assemblies, and even small intact cells to medium (∼4-8 nm) resolution in a near-native state, but restrictions in the range of accessible views are a major limitation. Here we report on the design, characterization, and demonstration of a new "flip-flop" rotation stage that allows facile and routine collection of two orthogonal tilt-series of cryosamples. Single- and dual-axis tomograms of a variety of samples are compared to illustrate qualitatively the improvement produced by inclusion of the second tilt-series. Exact quantitative expressions are derived for the volume of the remaining "missing pyramid" in reciprocal space. When orthogonal tilt-series are recorded to ±65° in each direction, as this new cryostage permits, only 11% of reciprocal space is left unmeasured. The tomograms suggest that further improvement could be realized, however, through better software to align and merge dual-axis tilt-series of cryosamples.

Original languageEnglish (US)
Pages (from-to)288-297
Number of pages10
JournalJournal of Structural Biology
Volume151
Issue number3
DOIs
Publication statusPublished - Sep 1 2005

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All Science Journal Classification (ASJC) codes

  • Structural Biology

Cite this

Iancu, C. V., Wright, E. R., Benjamin, J., Tivol, W. F., Dias, D. P., Murphy, G. E., ... Jensen, G. J. (2005). A "flip-flop" rotation stage for routine dual-axis electron cryotomography. Journal of Structural Biology, 151(3), 288-297. https://doi.org/10.1016/j.jsb.2005.07.004