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Related Concept Videos

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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.
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Overview of Electron Microscopy01:25

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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.
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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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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...
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Transmission Electron Microscopy01:15

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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...
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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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Volume scanning electron microscopy for imaging biological ultrastructure.

Benjamin Titze1, Christel Genoud2

  • 1Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland. benjamin.titze@fmi.ch.

Biology of the Cell
|July 20, 2016
PubMed
Summary

Advancements in volume electron microscopy (EM), particularly scanning EM (SEM), enable rapid, high-resolution 3D imaging of biological structures. This review details serial block-face SEM, focused ion beam SEM, and ATUM-SEM techniques for enhanced biological research.

Keywords:
Brain/nervous systemCellular imagingElectron microscopySystems biology

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Area of Science:

  • Biology
  • Microscopy
  • Biotechnology

Background:

  • Electron microscopy (EM) has been crucial for biological ultrastructure investigation for over 60 years.
  • Traditional EM methods for 3D imaging were manual, slow, and prone to errors.

Purpose of the Study:

  • To introduce and compare three novel volume scanning electron microscopy (SEM) techniques.
  • To provide an overview of the 3D dataset acquisition workflow using volume SEM.
  • To showcase diverse applications of volume SEM in biological research.

Main Methods:

  • Serial Block-Face Electron Microscopy (SBEM)
  • Focused Ion Beam SEM (FIB-SEM)
  • Automated Tape-Collecting Ultramicrotome SEM (ATUM-SEM)

Main Results:

  • Volume EM techniques, primarily SEM-based, offer faster and more reliable 3D imaging of cells and tissues.
  • Automated methods significantly improve z-resolution and reduce manual labor compared to traditional sectioning.
  • Software tools facilitate manipulation and quantitative analysis of 3D image stacks.

Conclusions:

  • Volume SEM methods revolutionize 3D ultrastructural analysis in biology.
  • These techniques provide high-resolution insights into cellular and tissue architecture.
  • Volume SEM is a powerful tool for diverse biological research applications.