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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...
Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

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.
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...

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Related Experiment Video

Updated: Jul 19, 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

Cryo-electron microscopy of biological samples.

M Joseph Costello1

  • 1Department of Cell and Developmental Biology, University of North Carolina at Chapel Hill, 27599, USA. mjc@med.unc.edu

Ultrastructural Pathology
|November 9, 2006
PubMed
Summary

Cryo-electron microscopy offers advanced methods for imaging biological tissues. Key techniques include frozen-hydrated samples for high-resolution protein structures and high-pressure freezing for detailed tissue ultrastructure and immunolabeling.

Area of Science:

  • Structural biology
  • Biophysics
  • Microscopy

Background:

  • Cryo-electron microscopy (cryo-EM) is crucial for visualizing biological structures at high resolution.
  • Integral membrane proteins and complex tissue architectures present unique challenges for imaging.
  • Advancements in sample preparation and imaging techniques are vital for unlocking cellular details.

Purpose of the Study:

  • To summarize current cryo-electron microscopy methods for biological tissue processing and imaging.
  • To highlight the importance of specific preparation techniques for different biological samples.
  • To introduce the potential of cryo-electron tomography for 3D ultrastructural analysis.

Main Methods:

  • Focus on frozen-hydrated sample preparation for cryo-electron crystallography, enabling near-atomic resolution of integral membrane proteins.

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

Published on: July 26, 2014

Preparing Lamellae from Vitreous Biological Samples Using a Dual-Beam Scanning Electron Microscope for Cryo-Electron Tomography
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Preparing Lamellae from Vitreous Biological Samples Using a Dual-Beam Scanning Electron Microscope for Cryo-Electron Tomography

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Last Updated: Jul 19, 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

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

Preparing Lamellae from Vitreous Biological Samples Using a Dual-Beam Scanning Electron Microscope for Cryo-Electron Tomography
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Preparing Lamellae from Vitreous Biological Samples Using a Dual-Beam Scanning Electron Microscope for Cryo-Electron Tomography

Published on: August 5, 2021

  • Detail high-pressure freezing for vitrifying large biological samples, suitable for ultrastructural and immunolabeling analysis.
  • Summarize methods for examining samples on a transmission electron microscope (TEM) cryo-stage, including cryo-electron tomography.

    • Main Results:

    • Frozen-hydrated samples facilitate high-resolution cryo-electron crystallography of membrane proteins.
    • High-pressure freezing effectively vitrifies diverse and large tissue samples for detailed ultrastructural studies.
    • Cryo-electron tomography provides a new avenue for visualizing the 3D structure of subcellular components.

    Conclusions:

    • Cryo-EM sample preparation methods like frozen-hydration and high-pressure freezing are essential for advancing structural biology.
    • These techniques enable detailed ultrastructural and immunolabeling analyses of various biological tissues.
    • Cryo-electron tomography holds significant promise for future 3D imaging of cellular components.