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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...

Preparation of Samples for Electron Microscopy

Preparation of Samples for Electron Microscopy

To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...

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Updated: Jun 29, 2025

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

Cryo-EM sample preparation for high-resolution structure studies.

Liguo Wang¹, Christina M Zimanyi²

¹Laboratory for BioMolecular Structure, Brookhaven National Laboratory, Upton, NY 11973, USA.

Acta Crystallographica. Section F, Structural Biology Communications

|March 26, 2024

Summary

This summary is machine-generated.

Single-particle cryo-electron microscopy (SPA cryo-EM) is a powerful tool for determining biomolecular structures. This guide offers practical advice on sample preparation to improve success rates for new SPA cryo-EM projects.

Keywords:

cryo-electron microscopy high-resolution structure determination single-particle analysis specimen preparation

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Last Updated: Jun 29, 2025

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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Preparation of High-Temperature Sample Grids for Cryo-EM

Preparation of High-Temperature Sample Grids for Cryo-EM

Published on: July 26, 2021

Area of Science:

Structural Biology
Biophysics
Biochemistry

Background:

Single-particle cryo-electron microscopy (SPA cryo-EM) is increasingly used for high-resolution biomolecular structure determination.
The growing adoption of SPA cryo-EM necessitates accessible guidance for researchers new to the technique.

Purpose of the Study:

To provide practical strategies and advice for sample preparation in SPA cryo-EM.
To highlight common challenges and pitfalls encountered during SPA cryo-EM sample preparation.
To enhance the success rate of new SPA cryo-EM projects.

Main Methods:

Review of common sample preparation techniques for SPA cryo-EM.
Identification of critical factors influencing sample quality for cryo-EM.
Discussion of potential pitfalls and troubleshooting strategies.

Main Results:

Outlines essential considerations for successful SPA cryo-EM sample preparation.
Identifies key strategies to optimize sample behavior and data quality.
Provides practical tips to navigate common challenges in the workflow.

Conclusions:

Effective sample preparation is crucial for successful SPA cryo-EM structure determination.
Understanding technique limitations and employing best practices increases project success.
This guide aims to empower researchers to confidently initiate SPA cryo-EM studies.