Three-Dimensional Reconstruction with Contrast Transfer Function Correction from Energy-Filtered Cryoelectron Micrographs: Procedure and Application to the 70S Escherichia coli Ribosome

Three-Dimensional Reconstruction with Contrast Transfer Function Correction from Energy-Filtered... Cryoelectron microscopy provides the means of studying macromolecules in their native state. However, the contrast transfer function (CTF) makes the images and the three-dimensional (3D) maps derived from them difficult to interpret. We developed methods to determine the CTF from experimental data and to obtain a CTF-corrected 3D reconstruction. The CTF correction and 3D reconstruction accomplished in one step make it easy to combine different defocus data sets and decrease the error accumulation in the computation. These methods were applied to energy-filtered images of the 70S Escherichia coli ribosome, resulting in a distortion-free 3D map of the ribosome at 1/24.5 Å −1 resolution, as determined by the differential phase residual resolution criterion. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Structural Biology Elsevier

Three-Dimensional Reconstruction with Contrast Transfer Function Correction from Energy-Filtered Cryoelectron Micrographs: Procedure and Application to the 70S Escherichia coli Ribosome

Journal of Structural Biology, Volume 118 (3) – Apr 1, 1997

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Publisher
Elsevier
Copyright
Copyright © 1997 Academic Press
ISSN
1047-8477
eISSN
1095-8657
DOI
10.1006/jsbi.1997.3845
Publisher site
See Article on Publisher Site

Abstract

Cryoelectron microscopy provides the means of studying macromolecules in their native state. However, the contrast transfer function (CTF) makes the images and the three-dimensional (3D) maps derived from them difficult to interpret. We developed methods to determine the CTF from experimental data and to obtain a CTF-corrected 3D reconstruction. The CTF correction and 3D reconstruction accomplished in one step make it easy to combine different defocus data sets and decrease the error accumulation in the computation. These methods were applied to energy-filtered images of the 70S Escherichia coli ribosome, resulting in a distortion-free 3D map of the ribosome at 1/24.5 Å −1 resolution, as determined by the differential phase residual resolution criterion.

Journal

Journal of Structural BiologyElsevier

Published: Apr 1, 1997

References

  • Three-dimensional reconstruction of the 70S Escherichia coli
    Frank, J.; Penczek, P.; Grassucci, R.; Srivastava, S.
  • SPIDER and WEB: Processing and visualization of images in 3D electron microscopy and related fields
    Frank, J.; Radermacher, M.; Penczek, P.; Zhu, J.; Li, Y.; Ladjadj, M.; Leith, A.
  • Electron microscopy of frozen biological suspensions
    Lepault, J.; Booy, F.P.; Dubochet, J.
  • A small particulate component of the cytoplasm
    Palade, G.E.
  • Components of bacterial ribosomes
    Wittmann, H.G.

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