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Related Concept Videos

Overview of Electron Microscopy01:25

Overview of Electron Microscopy

The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
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.
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Super-resolution Fluorescence Microscopy01:37

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Cryo-electron Microscopy01:28

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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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Updated: Jul 3, 2026

Cryo-EM and Single-Particle Analysis with Scipion
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Published on: May 29, 2021

Sharpening high resolution information in single particle electron cryomicroscopy.

J J Fernández1, D Luque, J R Castón

  • 1Centro Nacional de Biotecnologia, CSIC Campus Universidad Autonoma, Cantoblanco, 28049 Madrid, Spain. jj.fernandez@cnb.csic.es

Journal of Structural Biology
|July 11, 2008
PubMed
Summary

EM-BFACTOR software objectively determines B-factors to restore contrast in single-particle electron cryomicroscopy. This enhances visualization of high-resolution molecular features in experimental density maps for better interpretation.

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A Robust Single-Particle Cryo-Electron Microscopy (cryo-EM) Processing Workflow with cryoSPARC, RELION, and Scipion
13:43

A Robust Single-Particle Cryo-Electron Microscopy (cryo-EM) Processing Workflow with cryoSPARC, RELION, and Scipion

Published on: January 31, 2022

Area of Science:

  • Structural Biology
  • Biophysics
  • Microscopy

Background:

  • Single-particle electron cryomicroscopy (cryo-EM) achieves subnanometer resolution, revealing secondary structure elements.
  • Contrast loss in cryo-EM hinders interpretation of high-resolution structural features.
  • Traditional contrast loss modeling uses a Gaussian decay (B-factor), with restoration often relying on ad hoc methods.

Purpose of the Study:

  • To introduce and evaluate EM-BFACTOR, a program facilitating objective B-factor determination and contrast restoration.
  • To investigate a novel sharpening method for improving cryo-EM density maps.
  • To enhance the analysis of high-resolution cryo-EM data.

Main Methods:

  • Utilized the novel objective B-factor determination and contrast restoration method by Rosenthal and Henderson (2003).
  • Developed the EM-BFACTOR program to integrate with common cryo-EM software packages.
  • Investigated the sharpening method's effectiveness in unraveling concealed molecular features.

Main Results:

  • EM-BFACTOR program successfully facilitates objective B-factor determination and contrast restoration.
  • The investigated sharpening method effectively enhances the visibility of high-resolution molecular features.
  • Experimental density maps are improved, making them more amenable to interpretation.

Conclusions:

  • The EM-BFACTOR program and its associated sharpening method significantly improve the interpretation of cryo-EM data.
  • This approach aids in unraveling high-resolution molecular details previously obscured by contrast loss.
  • The method has the potential to facilitate routine analysis in high-resolution single-particle electron cryomicroscopy.