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Related Concept Videos

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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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
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Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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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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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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Related Experiment Video

Updated: May 4, 2026

Processing Embryo, Eggshell, and Fungal Culture for Scanning Electron Microscopy
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Metal shadowing for electron microscopy.

Gregory M Hendricks1

  • 1Core Electron Microscopy Facility, Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 21, 2013
PubMed
Summary

Metal shadowing offers high-resolution ultrastructure imaging for biological specimens like viruses and molecules. Advanced preparation techniques ensure superior preservation compared to air drying, preventing specimen crushing.

Area of Science:

  • Microscopy
  • Biophysics
  • Structural Biology

Background:

  • Metal shadowing is a high-resolution technique for visualizing biological ultrastructure.
  • Specimen preparation is critical for successful metal shadowing.
  • Air drying, used in negative staining, can damage delicate biological structures due to surface tension.

Purpose of the Study:

  • To detail specimen preparation techniques for metal shadowing.
  • To explain the development and procedures for preparing various biological samples.
  • To highlight the advantages of metal shadowing over air drying methods.

Main Methods:

  • Evaporation of heavy metals at an oblique angle to the specimen.
  • Development of complex specimen preparation techniques preceding metal deposition.

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  • A variation involves rotating the specimen during metal deposition for 3D rendering.

    • Main Results:

    • Metal shadowing creates a "shadow" effect, revealing surface topography.
    • Pre-shadowing preparation techniques provide superior preservation of biological specimens.
    • The method is applicable to bacteria, viruses, isolated molecules, and macromolecular assemblies.

    Conclusions:

    • Metal shadowing is a powerful technique for ultrastructural analysis of diverse biological samples.
    • Careful specimen preparation is essential for optimal results and preservation.
    • 3D rendering is possible with specimen rotation during metal deposition.