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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...
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: May 16, 2026

Single Particle Cryo-Electron Microscopy: From Sample to Structure
11:52

Single Particle Cryo-Electron Microscopy: From Sample to Structure

Published on: May 29, 2021

In situ single-particle Cryo-EM methods: Principle and applications.

Xing Zhang1, Guoqiang Huang1, Sen-Fang Sui2

  • 1State Key Laboratory of Membrane Biology, Beijing Frontier Research Center for Biological Structures, School of Life Sciences, Tsinghua University, Beijing, China.

Current Opinion in Structural Biology
|May 14, 2026
PubMed
Summary
This summary is machine-generated.

Cryo-electron microscopy (cryo-EM) now visualizes cellular structures in situ. New single-particle cryo-EM methods overcome resolution limits of older techniques, improving macromolecular architecture analysis.

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Related Experiment Videos

Last Updated: May 16, 2026

Single Particle Cryo-Electron Microscopy: From Sample to Structure
11:52

Single Particle Cryo-Electron Microscopy: From Sample to Structure

Published on: May 29, 2021

Cryo-EM and Single-Particle Analysis with Scipion
09:06

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Published on: May 29, 2021

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

Area of Science:

  • Structural Biology
  • Cellular Imaging
  • Biophysics

Background:

  • Cryo-electron microscopy (cryo-EM) is advancing from in vitro protein analysis to in situ cellular visualization.
  • Conventional cryo-electron tomography (cryo-ET) with subtomogram averaging faces resolution limitations due to complex workflows and low throughput.
  • Emerging in situ single-particle cryo-EM offers a promising alternative for high-resolution cellular imaging.

Purpose of the Study:

  • To review the principles, workflows, and advantages of in situ single-particle cryo-EM methods.
  • To highlight key applications of these advanced cryo-EM techniques.
  • To discuss future perspectives in in situ structural biology.

Main Methods:

  • Collecting high-dose, untilted images of cellular lamellae using in situ single-particle cryo-EM.
  • Utilizing high-resolution templates for particle identification and refinement.
  • Overcoming limitations of conventional cryo-electron tomography (cryo-ET) and subtomogram averaging.

Main Results:

  • Significant advancements in data throughput for in situ cryo-EM.
  • Substantial improvements in achievable resolution for macromolecular architecture.
  • Enabling direct visualization of cellular structures with unprecedented detail.

Conclusions:

  • In situ single-particle cryo-EM methods represent a major leap in visualizing cellular architecture.
  • These methods address key limitations of traditional cryo-ET, offering higher resolution and throughput.
  • The review provides a comprehensive overview and outlook for this rapidly evolving field.