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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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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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Electron Microscope Tomography and Single-particle Reconstruction01:07

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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
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Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

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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...
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Related Experiment Video

Updated: Mar 7, 2026

Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction
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Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction

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Structural Study of Heterogeneous Biological Samples by Cryoelectron Microscopy and Image Processing.

H E White1, A Ignatiou1, D K Clare1

  • 1Institute of Structural and Molecular Biology, University College London and Birkbeck, Malet Street, London WC1E 7HX, UK.

Biomed Research International
|February 14, 2017
PubMed
Summary
This summary is machine-generated.

Modern electron microscopy, including single particle analysis (SPA), visualizes biological macromolecules in multiple states. This review details techniques for separating conformational states to reveal complex biological mechanisms.

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Single Particle Cryo-Electron Microscopy: From Sample to Structure
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Area of Science:

  • Structural biology
  • Biophysics
  • Biochemistry

Background:

  • Biological macromolecules exhibit intrinsic flexibility and exist in multiple conformations.
  • Modern electron microscopy, particularly cryo-EM, visualizes biocomplexes in near-native states.
  • Advances in technology and software have enhanced EM resolution and revealed structural variations.

Purpose of the Study:

  • To review principles of techniques used for analyzing structural conformations.
  • To provide examples of successful applications in studying biologically significant complexes.
  • To highlight methods for separating different conformational states.

Main Methods:

  • Single particle analysis (SPA) for separating conformational states.
  • 2D and 3D image analysis techniques.
  • Computational approaches for large datasets and high-resolution structure determination.

Main Results:

  • SPA offers approaches to separate distinct conformational states of biocomplexes.
  • Advanced EM techniques reveal structural variations and improve resolution.
  • Successful applications demonstrate the utility of these methods in structural biology.

Conclusions:

  • Understanding conformational dynamics is crucial for elucidating biological mechanisms.
  • Cryo-EM and SPA are powerful tools for high-resolution structural studies.
  • The reviewed techniques enable detailed analysis of macromolecular flexibility and function.