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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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Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
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Related Experiment Video

Updated: Mar 12, 2026

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE

Published on: May 16, 2017

Antibody-Based Affinity Cryoelectron Microscopy at 2.6-Å Resolution.

Guimei Yu¹, Kunpeng Li¹, Pengwei Huang²

¹Department of Biological Science, Markey Center for Structural Biology, Purdue University, West Lafayette, IN 47907, USA.

Structure (London, England : 1993)

|November 3, 2016

Summary

Affinity cryo-electron microscopy (cryo-EM) now achieves near-atomic resolution for challenging samples. This antibody-based method enables high-resolution structural analysis of low-abundance proteins previously out of reach for cryo-EM.

Keywords:

Tulane virus affinity cryoelectron microscopy antibody-based affinity grid single-particle 3D reconstruction

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Last Updated: Mar 12, 2026

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE

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Published on: March 16, 2022

Single Particle Cryo-Electron Microscopy: From Sample to Structure

Single Particle Cryo-Electron Microscopy: From Sample to Structure

Published on: May 29, 2021

Area of Science:

Structural Biology
Biophysics
Microscopy

Background:

Affinity cryo-electron microscopy (cryo-EM) simplifies sample preparation for cryo-EM.
This technique aims to analyze challenging specimens, including those of low abundance or unpurified.
Current limitations include the inability to achieve near-atomic resolution, leading to a perception of it as a niche method.

Purpose of the Study:

To demonstrate that affinity cryo-EM can achieve near-atomic resolution.
To solve the structure of a challenging, low-abundance sample using affinity cryo-EM.
To compare the quality of affinity cryo-EM with conventional methods.

Main Methods:

Utilized an antibody-based affinity grid approach for sample preparation.
Applied cryo-EM to analyze low-concentration Tulane virus from a low-yield cell-culture system.
Performed quantitative analyses to assess data and reconstruction quality.

Main Results:

Successfully solved a ∼2.6-Å structure of Tulane virus.
The sample was challenging due to low concentration and low yield, making it difficult for standard cryo-EM.
Quantitative analyses showed data and reconstruction quality comparable to conventional methods using high-concentration samples.

Conclusions:

Affinity cryo-EM is capable of achieving near-atomic resolution, even for difficult samples.
The antibody-based affinity grid approach overcomes limitations of standard cryo-EM for low-abundance specimens.
This advancement expands the utility of cryo-EM for structural biology research.