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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...
Transmission Electron Microscopy01:15

Transmission Electron Microscopy

In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400 keV in...
Scanning Electron Microscopy01:07

Scanning Electron Microscopy

A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a two-dimensional manner. Typically, areas between ~1 centimeter to 5 micrometers in width can be imaged. SEM can be used to image bacteria, viruses, tissues as well as larger samples like insects. Conventional SEM gives a magnification ranging from 20X to 30,000X and spatial resolution of 50 to 100 nanometers.
Fundamental Principles
Accelerated...
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.
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
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...

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Related Experiment Video

Updated: Jul 9, 2026

Array Tomography Workflow for the Targeted Acquisition of Volume Information using Scanning Electron Microscopy
09:47

Array Tomography Workflow for the Targeted Acquisition of Volume Information using Scanning Electron Microscopy

Published on: July 15, 2021

Automated electron tomography with scanning transmission electron microscopy.

Jianglin Feng1, Andrew P Somlyo, Avril V Somlyo

  • 1Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA 22908, USA. jf4x@virginia.edu

Journal of Microscopy
|November 30, 2007
PubMed
Summary

A new automated system for scanning transmission electron microscopy (STEM) tomography enables high-resolution 3D imaging. This advanced technique corrects specimen movement and tilt for clearer reconstructions of large objects.

More Related Videos

Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography
08:04

Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography

Published on: March 12, 2017

Related Experiment Videos

Last Updated: Jul 9, 2026

Array Tomography Workflow for the Targeted Acquisition of Volume Information using Scanning Electron Microscopy
09:47

Array Tomography Workflow for the Targeted Acquisition of Volume Information using Scanning Electron Microscopy

Published on: July 15, 2021

Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography
08:04

Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography

Published on: March 12, 2017

Area of Science:

  • Electron Microscopy
  • Materials Science
  • Nanotechnology

Background:

  • Automated data collection is crucial for advancing 3D imaging techniques.
  • Scanning Transmission Electron Microscopy (STEM) offers high resolution but requires precise sample manipulation.
  • Tomography provides 3D structural information from 2D projection images.

Purpose of the Study:

  • To develop and implement a fully automated data collection system for STEM tomography.
  • To improve the accuracy and efficiency of 3D reconstructions in electron microscopy.
  • To overcome limitations of traditional TEM tomography for large object analysis.

Main Methods:

  • Implemented an automated system combining mechanical and electronic corrections for specimen autotracking.
  • Utilized contrast difference in focus series for an automated autofocusing routine.
  • Employed dynamic focusing to eliminate focus gradients caused by specimen tilt.

Main Results:

  • Successfully demonstrated a fully automated tomographic data collection system in STEM mode.
  • Autotracking and autofocusing mechanisms effectively corrected for specimen movement and tilt.
  • Dynamic focusing successfully removed focus gradients, enhancing image quality.

Conclusions:

  • The automated STEM tomography system enables efficient and high-resolution 3D imaging.
  • This method offers advantages over TEM tomography, particularly for reconstructing large objects.
  • The developed system has the potential to significantly advance structural analysis in materials science and nanotechnology.