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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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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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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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A Device for Ribbon Collection for Array Tomography with Scanning Electron Microscopy.

Taro Koike1, Yosky Kataoka2,3, Mitsuyo Maeda2

  • 1Department of Anatomy and Cell Science, Kansai Medical University, Hirakata City, Osaka, Japan.

Acta Histochemica Et Cytochemica
|December 26, 2017
PubMed
Summary

Researchers developed an inexpensive device for array tomography, simplifying the mounting of numerous ultrathin sections for 3D cell and tissue imaging using scanning electron microscopy (SEM). This method enhances 3D ultrastructural analysis.

Keywords:
array tomographypost-embedding labelingscanning electron microscopyserial ultrathin sectionthree-dimensional observation

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biotechnology

Background:

  • Array tomography enables 3D visualization of cellular and tissue fine structure.
  • Manual mounting of serial ultrathin sections for array tomography is challenging and labor-intensive.
  • Existing methods limit the number of sections that can be efficiently mounted on substrates.

Purpose of the Study:

  • To develop an inexpensive, laboratory-made device to automate the mounting of serial ultrathin sections for array tomography.
  • To improve the efficiency and reproducibility of preparing samples for scanning electron microscopy (SEM).
  • To facilitate the use of array tomography with post-embedding immunocytochemistry.

Main Methods:

  • A novel device was engineered using a nylon fishing line to lift a substrate from a knife boat, facilitating ribbon mounting.
  • The device was tested for its ability to mount multiple serial ultrathin sections (ribbons).
  • Specimens were prepared using water-soluble resin blocks and subjected to post-embedding immunocytochemistry.

Main Results:

  • The device successfully mounted an average of 205.6 serial ultrathin sections per substrate.
  • The method proved effective for ribbons derived from water-soluble resin blocks.
  • Successful staining via post-embedding immunocytochemistry confirmed the method's compatibility with biological labeling.

Conclusions:

  • The developed laboratory-made device significantly simplifies and enhances the mounting process for array tomography.
  • This cost-effective method increases the number of sections mountable, improving 3D ultrastructural analysis via SEM.
  • The technique is suitable for various sample types and compatible with immunocytochemistry, broadening its applicability in biological research.