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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: Jun 1, 2026

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

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

Published on: February 7, 2022

Limiting factors in atomic resolution cryo electron microscopy: no simple tricks.

Xing Zhang¹, Z Hong Zhou

¹Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, 237 BSRB, 615 Charles E. Young Dr. S., Los Angeles, CA 90095-7364, USA.

Journal of Structural Biology

|June 2, 2011

Summary

Achieving atomic resolution in cryo electron microscopy (cryoEM) requires addressing electron beam tilt, defocus inaccuracies, focus gradients, and dynamic scattering. Strategies are proposed to overcome these limitations for improved 3D reconstructions.

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Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction

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Published on: January 9, 2015

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Last Updated: Jun 1, 2026

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

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

Published on: February 7, 2022

Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction

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

Area of Science:

Structural Biology
Biophysics
Microscopy

Background:

Cryo electron microscopy (cryoEM) is crucial for determining the structure of large biological complexes.
Achieving atomic resolution with cryoEM is challenging due to several limiting factors in image acquisition and 3D reconstruction.

Purpose of the Study:

To analyze key factors limiting atomic resolution in cryoEM.
To propose strategies for overcoming these limitations in cryoEM image acquisition and 3D reconstruction.

Main Methods:

Analysis of electron beam tilt, defocus determination, focus gradients, and dynamic electron scattering.
Development of strategies including parallel illumination, coma-free alignment, spherical aberration correction, dose-dependent imaging, iterative model-based methods, and deconvolution techniques.

Main Results:

Identified electron beam tilt, defocus inaccuracies, focus gradients, and dynamic scattering as significant limitations.
Proposed specific methods to mitigate the impact of each identified factor on cryoEM data.

Conclusions:

The proposed strategies offer practical guidance for experimental design in cryoEM.
Addressing these factors is essential for advancing cryoEM towards routine atomic resolution reconstructions.