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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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Heating a crystalline solid increases the average energy of its atoms, molecules, or ions, and the solid gets hotter. At some point, the added energy becomes large enough to partially overcome the forces holding the molecules or ions of the solid in their fixed positions, and the solid begins the process of transitioning to the liquid state or melting. At this point, the temperature of the solid stops rising, despite the continual input of heat, and it remains constant until all of the solid is...

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Related Experiment Video

Updated: Mar 30, 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

A Simple Method for Quick-Freezing.

Elaine L Bearer¹, Lelio Orci¹

¹Department of Pathology, University of California, San Francisco, CA 94143 (E.L.B.); Institute d'Histologie, Department de Morphologie, Centre Medicale Universitaire, Geneva 1211, Switzerland (L.O.).

Journal of Electron Microscopy Technique

|November 10, 2015

Summary

This summary is machine-generated.

A new method for quick-freezing tissue without glycerol cryoprotection enables detailed visualization of cellular structures. This technique overcomes ice crystal formation, preserving molecular detail for freeze-fracture electron microscopy.

Keywords:

Balzers’ freeze fracture device Deep-etching Quick-freezing

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

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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

Related Experiment Videos

Last Updated: Mar 30, 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:

Electron Microscopy
Cell Biology
Biophysics

Background:

Conventional freeze-fracture microscopy requires glycerol cryoprotection, which hinders ice removal and obscures hydrophilic structures.
Glycerol-free cryoprotection methods face challenges with ice crystal formation, limiting preservation of fine structural detail.
Existing quick-freezing techniques may not adequately preserve molecular details for subsequent analysis.

Purpose of the Study:

To develop a simple, glycerol-free quick-freezing method for tissue samples.
To assess the effectiveness of this method in preserving molecular and cellular structures for freeze-fracture electron microscopy.
To compare the results with conventional cryoprotection and liquid helium-temperature freezing.

Main Methods:

A novel method for quick-freezing tissue samples on a nitrogen-cooled copper block using a hand-held specimen holder.
Elimination of glycerol cryoprotection to allow for subsequent ice etching or freeze-drying.
Analysis of preserved molecular and cellular structures using freeze-fracture electron microscopy.

Main Results:

The developed method effectively minimizes ice crystal formation in glycerol-free frozen tissue.
High-resolution visualization of molecular details in isolated proteins (ferritin, catalase) was achieved.
In situ cellular structures were well-preserved and visualized, comparable to or exceeding other methods.

Conclusions:

This simple, glycerol-free quick-freezing technique provides excellent preservation of fine structural detail for freeze-fracture electron microscopy.
The method facilitates the visualization of both molecular components and intricate cellular architecture.
It offers a viable alternative to glycerol-based cryoprotection, enhancing the study of biological structures.