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Related Concept Videos

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...
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Molecular Models02:00

Molecular Models

Physical models representing molecular architectures of chemical compounds play essential roles in understanding chemistry. The use of molecular models makes it easier to visualize the structures and shapes of atoms and molecules.

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Related Experiment Video

Updated: Jun 6, 2026

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE
13:28

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE

Published on: May 16, 2017

Visualizing molecular machines in action: Single-particle analysis with structural variability.

Sjors H W Scheres1

  • 1MRC Laboratory of Molecular Biology, Cambridge, United Kingdom.

Advances in Protein Chemistry and Structural Biology
|December 1, 2010
PubMed
Summary
This summary is machine-generated.

Structural variability in electron microscopy (EM) samples, once a challenge, can now be leveraged. Advanced image processing allows classifying different molecular states for multiple 3D reconstructions from single datasets.

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Visualizing Single Molecular Complexes In Vivo Using Advanced Fluorescence Microscopy
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Published on: September 8, 2009

Related Experiment Videos

Last Updated: Jun 6, 2026

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE
13:28

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE

Published on: May 16, 2017

Visualizing Single Molecular Complexes In Vivo Using Advanced Fluorescence Microscopy
11:26

Visualizing Single Molecular Complexes In Vivo Using Advanced Fluorescence Microscopy

Published on: September 8, 2009

Area of Science:

  • Biophysics
  • Structural Biology
  • Biochemistry

Background:

  • Macromolecular assemblies are often flexible, existing in multiple structural states during function.
  • This structural heterogeneity in electron microscopy (EM) samples complicates single-particle analysis (SPA) and can lead to reconstruction artifacts.

Purpose of the Study:

  • To review recent advancements in image processing for single-particle analysis.
  • To demonstrate how structural variability can be transformed from a hindrance into an advantage for studying molecular dynamics.

Main Methods:

  • Classification algorithms for sorting projection images based on their 3D structures.
  • Single-particle analysis (SPA) techniques applied to heterogeneous EM datasets.

Main Results:

  • Modern algorithms enable the classification of projection images, allowing for the resolution of multiple 3D structures from a single dataset.
  • Structural variability can be effectively managed and utilized in 3D-EM.

Conclusions:

  • 3D Electron Microscopy (3D-EM) is uniquely positioned to study the dynamic behavior of molecular assemblies.
  • Advancements in image processing have turned structural heterogeneity into a powerful tool for structural biology research.