Practical Experience with Hole-Free Phase Plates for Cryo Electron Microscopy

Overview

Journal Microsc Microanal

Publisher Oxford University Press

Date 2016 Nov 25

PMID 27881198

Citations 2

Authors

Michael Marko

Chyongere Hsieh

Eric Leith

David Mastronarde

Sohei Motoki

Affiliations

Soon will be listed here.

Abstract

Phase plate (PP) imaging has proven to be valuable in transmission cryo electron microscopy of unstained, native-state biological specimens. Many PP types have been described, however until the recent implementation of the "hole-free" phase plate (HFPP), imaging has been challenging. We found the HFPP to be simple to construct and to set up in the transmission electron microscopy, but care in implementing automated data collection is needed. Performance may be variable, both initially and over time, thus it is important to monitor and evaluate image quality by observing the power spectrum. We found that while some HFPPs gave transfer to high resolution without CTF oscillation, most reached high resolution when operated with modest defocus.

Citing Articles

Analyzing contrast in cryo-transmission electron microscopy: Comparison of electrostatic Zach phase plates and hole-free phase plates.

Obermair M, Hettler S, Hsieh C, Dries M, Marko M, Gerthsen D Ultramicroscopy. 2020; 218:113086.

PMID: 32781400 PMC: 8848455. DOI: 10.1016/j.ultramic.2020.113086.

In situ Microfluidic Cryofixation for Cryo Focused Ion Beam Milling and Cryo Electron Tomography.

Fuest M, Schaffer M, Nocera G, Galilea-Kleinsteuber R, Messling J, Heymann M Sci Rep. 2019; 9(1):19133.

PMID: 31836773 PMC: 6911106. DOI: 10.1038/s41598-019-55413-2.

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