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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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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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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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Scanning Electron Microscopy01:07

Scanning Electron Microscopy

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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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Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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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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Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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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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Related Experiment Video

Updated: Feb 1, 2026

Sample Preparation and Imaging of Exosomes by Transmission Electron Microscopy
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Sample Preparation for Transmission Electron Microscopy.

Parastou Tizro1, Cecilia Choi1, Negar Khanlou2

  • 1Division of Neuropathology, Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 13, 2018
PubMed
Summary

Transmission electron microscopy (TEM) requires intricate sample preparation to withstand harsh microscope conditions. Proper techniques are crucial for preserving the native structure of biological materials during TEM analysis.

Keywords:
FixationHigh pressure freezingProtocolSample preparationTEM

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Area of Science:

  • Materials Science
  • Biological Sciences
  • Microscopy

Background:

  • Transmission electron microscopy (TEM) is vital for studying cellular and biological material structures.
  • Harsh conditions within electron microscopes (electron bombardment, vacuum evaporation) damage samples.
  • Complex sample preparation is essential to mitigate damage and enable electron transmission.

Purpose of the Study:

  • To detail the critical steps involved in preparing samples for Transmission Electron Microscopy.
  • To emphasize the importance of meticulous execution in each preparation stage.
  • To ensure the preservation of native sample structures during the preparation process.

Main Methods:

  • Specimen stabilization and reduction to small, thin sections (approx. 3 mm diameter, 300-500 nm thickness for biological studies).
  • Application of diverse preparation techniques tailored to specific studies and specimens.
  • Careful execution of each step to maintain sample integrity.

Main Results:

  • Preparation methods are designed to stabilize specimens for the electron microscope environment.
  • Achieving precise dimensions (thickness, diameter) is critical for electron transmission.
  • The ultimate goal is to preserve the native structure of the biological sample.

Conclusions:

  • Successful TEM analysis hinges on rigorous and detailed sample preparation.
  • Each preparation step significantly influences the final quality of the TEM data.
  • Adherence to precise protocols is paramount for accurate structural analysis of biological materials.