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

Research output: Contribution to journalArticlepeer-review

55 Scopus citations

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
StatePublished - Sep 2005

All Science Journal Classification (ASJC) codes

  • Structural Biology

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