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Cryo-electron Microscopy

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

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Related Experiment Video

Updated: Mar 11, 2026

Routine Collection of High-Resolution cryo-EM Datasets Using 200 KV Transmission Electron Microscope

Routine Collection of High-Resolution cryo-EM Datasets Using 200 KV Transmission Electron Microscope

Published on: March 16, 2022

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

Michael Marko¹, Chyongere Hsieh¹, Eric Leith²

¹1NY State Department of Health,Wadsworth Center,PO Box 509,Albany,NY 12201,USA.

Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada

|November 25, 2016

Summary

The hole-free phase plate (HFPP) simplifies transmission cryo-electron microscopy for biological samples. Careful setup and monitoring are crucial for optimal high-resolution imaging performance.

Keywords:

SerialEM contrast-transfer function cryo electron microscopy (cryo-EM)cryo electron tomography (cryo-ET)phase plate

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Last Updated: Mar 11, 2026

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Published on: March 16, 2022

Manual Blot-and-Plunge Freezing of Biological Specimens for Single-Particle Cryogenic Electron Microscopy

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Published on: February 7, 2022

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Area of Science:

Cryo-electron microscopy
Phase contrast imaging
Structural biology

Background:

Phase plate (PP) imaging is essential for visualizing unstained, native biological specimens in transmission cryo-electron microscopy (cryo-EM).
Previous phase plate designs presented challenges in practical implementation and data acquisition.

Purpose of the Study:

To evaluate the utility and performance of the hole-free phase plate (HFPP) in transmission cryo-EM.
To provide guidance on the setup and data collection using HFPPs for optimal imaging.

Main Methods:

Construction and implementation of the hole-free phase plate (HFPP) in a transmission electron microscope.
Automated data collection strategies were employed.
Image quality was assessed by analyzing the power spectrum and monitoring for contrast transfer function (CTF) oscillations.

Main Results:

The HFPP is straightforward to construct and integrate into transmission electron microscopy workflows.
Automated data collection requires careful implementation to ensure consistent performance.
While some HFPPs achieved high-resolution transfer without CTF oscillations, most performed optimally with modest defocus.
Performance can vary over time, necessitating regular image quality evaluation.

Conclusions:

The hole-free phase plate (HFPP) offers a valuable advancement for cryo-EM imaging of biological samples.
Successful application of HFPPs requires meticulous attention to setup, data collection parameters, and ongoing performance monitoring.