Cryo-EM single particle analysis with the Volta phase plate
Abstract
We present a method for in-focus data acquisition with a phase plate that enables near-atomic resolution single particle reconstructions. Accurate focusing is the determining factor for obtaining high quality data. A double-area focusing strategy was implemented in order to achieve the required precision. With this approach we obtained a 3.2 Å resolution reconstruction of the Thermoplasma acidophilum 20S proteasome. The phase plate matches or slightly exceeds the performance of the conventional defocus approach. Spherical aberration becomes a limiting factor for achieving resolutions below 3 Å with in-focus phase plate images. The phase plate could enable single particle analysis of challenging samples in terms of small size, heterogeneity and flexibility that are difficult to solve by the conventional defocus approach.
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Reviewing Editor
- Sjors HW Scheres, Medical Research Council, United Kingdom
Version history
- Received: November 15, 2015
- Accepted: March 3, 2016
- Accepted Manuscript published: March 7, 2016 (version 1)
- Version of Record published: April 28, 2016 (version 2)
Copyright
© 2016, Danev & Baumeister
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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