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Related Concept Videos

Cryo-electron Microscopy01:28

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: Jun 8, 2026

Manual Blot-and-Plunge Freezing of Biological Specimens for Single-Particle Cryogenic Electron Microscopy
09:16

Manual Blot-and-Plunge Freezing of Biological Specimens for Single-Particle Cryogenic Electron Microscopy

Published on: February 7, 2022

Plunge freezing for electron cryomicroscopy.

Megan J Dobro1, Linda A Melanson, Grant J Jensen

  • 1Division of Biology, California Institute of Technology, Pasadena, California, USA.

Methods in Enzymology
|October 5, 2010
PubMed
Summary

Plunge freezing is the preferred method for preserving aqueous biological samples for electron microscopy, ensuring they remain in a native state. This technique requires careful control of sample preparation, cryogen, and plunging conditions for optimal results.

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

Published on: May 5, 2017

Cryo-electron Microscopy Specimen Preparation By Means Of a Focused Ion Beam
10:54

Cryo-electron Microscopy Specimen Preparation By Means Of a Focused Ion Beam

Published on: July 26, 2014

Related Experiment Videos

Last Updated: Jun 8, 2026

Manual Blot-and-Plunge Freezing of Biological Specimens for Single-Particle Cryogenic Electron Microscopy
09:16

Manual Blot-and-Plunge Freezing of Biological Specimens for Single-Particle Cryogenic Electron Microscopy

Published on: February 7, 2022

Plunge Freezing: A Tool for the Ultrastructural and Immunolocalization Studies of Suspension Cells in Transmission Electron Microscopy
13:35

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

Published on: May 5, 2017

Cryo-electron Microscopy Specimen Preparation By Means Of a Focused Ion Beam
10:54

Cryo-electron Microscopy Specimen Preparation By Means Of a Focused Ion Beam

Published on: July 26, 2014

Area of Science:

  • Electron Microscopy
  • Biophysics
  • Cryo-EM Sample Preparation

Background:

  • Aqueous biological samples require stabilization for high-vacuum electron microscopy.
  • Plunge freezing is a crucial technique for preserving sample integrity in its native state.
  • Successful cryo-electron microscopy (cryo-EM) relies on effective sample preservation.

Purpose of the Study:

  • To review the principles and techniques of plunge freezing for biological samples.
  • To discuss instrumentation, common challenges, and safety protocols associated with plunge freezing.
  • To highlight plunge freezing as the method of choice for native-state sample preservation in electron cryomicroscopy.

Main Methods:

  • Spreading the biological sample into a thin film on an electron microscopy (EM) grid.
  • Rapidly submerging the grid into a cryogen, typically liquid ethane.
  • Controlling critical parameters such as grid properties, sample film uniformity, cryogen characteristics, and plunging dynamics.

Main Results:

  • Plunge freezing, when optimized, successfully preserves biological samples in a near-native state.
  • Success is contingent upon meticulous control over sample application, cryogen properties, and plunging parameters.
  • This method is essential for high-resolution imaging in electron cryomicroscopy.

Conclusions:

  • Plunge freezing is a vital technique for preparing aqueous biological samples for electron microscopy.
  • Understanding the interplay of various factors is key to achieving high-quality, vitrified samples.
  • Adherence to established principles and safety guidelines ensures reliable and reproducible results in cryo-EM.