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Updated: Aug 10, 2026

Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction
Published on: January 9, 2015
The ribosome at improved resolution: new techniques for merging and orientation refinement in 3D cryo-electron
P A Penczek1, R A Grassucci, J Frank
1Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
Abstract:
Cryo-electron microscopy of single biological particles opens up new possibilities for structure analysis: the particle can be reconstructed in its native shape and internal features are preserved. To take advantage of these possibilities we have developed new methods of data collection and image processing and we have applied them to the 70S Escherichia coli ribosome. A method of orientation search is proposed, which makes it possible to relate random-conical data sets to one another even if they are collected from low-tilt micrographs. A technique for 3D alignment of projections is described and applied to the single-micrograph 3D reconstruction.
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