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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: May 4, 2026

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

Published on: October 13, 2014

Cryo-fixation by self-pressurized rapid freezing.

Markus Grabenbauer¹, Hong-Mei Han, Jan Huebinger

¹Institute for Anatomy and Cell Biology Germany, University of Heidelberg, Heidelberg, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|December 21, 2013

Summary

This summary is machine-generated.

Self-pressurized rapid freezing offers a low-cost, effective alternative to high-pressure freezing for preserving cellular ultrastructure. This method achieves sample quality comparable to traditional high-pressure freezing (HPF) for cryo-electron microscopy applications.

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Last Updated: May 4, 2026

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

Published on: October 13, 2014

Rapid Freezing using Sandwich Freezing Device for Good Ultrastructural Preservation of Biological Specimens in Electron Microscopy

Rapid Freezing using Sandwich Freezing Device for Good Ultrastructural Preservation of Biological Specimens in Electron Microscopy

Published on: July 19, 2021

Plunge Freezing: A Tool for the Ultrastructural and Immunolocalization Studies of Suspension Cells in Transmission Electron Microscopy

Plunge Freezing: A Tool for the Ultrastructural and Immunolocalization Studies of Suspension Cells in Transmission Electron Microscopy

Published on: May 5, 2017

Area of Science:

Cell Biology
Biophysics
Microscopy Techniques

Background:

High-pressure freeze fixation (HPF) is crucial for preserving cellular ultrastructure by instantly halting biological processes.
Vitrified samples from HPF can be analyzed using cryo-electron microscopy of vitreous sections (CEMOVIS) or processed via freeze substitution.
Achieving near-native states requires rapid and effective fixation methods.

Purpose of the Study:

To introduce a novel, cost-effective, and user-friendly freeze fixation technique.
To demonstrate that self-pressurized rapid freezing can yield results comparable to conventional HPF.
To provide an alternative method for preparing biological samples for high-resolution imaging and analysis.

Main Methods:

Samples (cells, small organisms) are placed in sealed capillary metal tubes.
Tubes are plunged into liquid ethane cooled by liquid nitrogen.
Crystalline ice formation within the tube generates pressure, inducing vitrification of the specimen.

Main Results:

The self-pressurized rapid freezing method effectively arrests dynamic processes in biological samples.
Sample quality achieved is equivalent to that obtained with conventional high-pressure freezing apparatus.
The method allows for high-resolution cryo-electron microscopy and subsequent processing like freeze substitution.

Conclusions:

Self-pressurized rapid freezing is a viable, accessible alternative to HPF for biological sample preparation.
This technique enables high-quality cryo-EM and correlative light and electron microscopy studies.
It expands the accessibility of advanced microscopy techniques for research.