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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 26, 2026

Author Spotlight: Enhancing Cryo-EM Sample Preparation with Streptavidin-Biotin Approach

Author Spotlight: Enhancing Cryo-EM Sample Preparation with Streptavidin-Biotin Approach

Published on: December 29, 2023

Antibody-based affinity cryo-EM grid.

Guimei Yu¹, Kunpeng Li¹, Wen Jiang¹

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

Methods (San Diego, Calif.)

|January 26, 2016

Summary

This summary is machine-generated.

The new cryo-SPIEM method simplifies cryo-electron microscopy (cryo-EM) grid preparation by integrating sample purification. This antibody-based approach enables direct study of native macromolecular complexes, including those in low abundance or transient states.

Keywords:

3D reconstruction Affinity grid Antibodies Cryo-SPIEM On-grid assays Single particle cryo-EM

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Published on: February 7, 2022

Area of Science:

Structural biology
Biophysics
Biochemistry

Background:

The Affinity Grid technique integrates sample purification and cryo-electron microscopy (cryo-EM) grid preparation into a single step.
Previous methods include functionalized lipid monolayers, streptavidin 2D crystals, and covalently functionalized carbon surfaces.
These methods often require target modifications or complex procedures.

Purpose of the Study:

To present a novel affinity cryo-EM approach, cryo-SPIEM (cryo-Solid Phase Immune Electron Microscopy).
To simplify affinity grid preparation for cryo-EM.
To enable direct study of native macromolecular complexes without target modification.

Main Methods:

Application of the traditional Solid Phase Immune Electron Microscopy (SPIEM) technique to cryo-EM.
Utilizing antibody-based affinity grids.
Direct analysis of native macromolecular complexes.

Main Results:

Cryo-SPIEM significantly simplifies the preparation of affinity grids.
The method directly works with native macromolecular complexes.
Enables cryo-EM studies of samples from cell cultures, low-abundance targets, and unstable intermediate states.

Conclusions:

The antibody-based affinity grid approach, cryo-SPIEM, offers a streamlined method for cryo-EM sample preparation.
This technique facilitates the investigation of previously challenging biological samples in their native states.
Broad applicability for studying diverse macromolecular complexes with high affinity and specificity antibodies.