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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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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 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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Biological Sample Preparation by High-pressure Freezing, Microwave-assisted Contrast Enhancement, and Minimal Resin Embedding for Volume Imaging
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Microwave-assisted processing and embedding for transmission electron microscopy.

Paul Webster1

  • 1Center for Electron Microscopy and Microanalysis (CEMMA), University of Souther California, Los Angeles, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 21, 2013
PubMed
Summary
This summary is machine-generated.

Rapid microwave processing significantly accelerates the preparation of biological samples for transmission electron microscopy. This method enables faster dehydration and resin embedding, reducing processing time to under 4 hours.

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Area of Science:

  • Biological Sciences
  • Microscopy Techniques
  • Materials Science

Background:

  • Transmission electron microscopy (TEM) requires meticulously prepared biological specimens.
  • Traditional methods for processing and embedding biological samples are time-consuming.
  • Rapid processing is crucial for time-sensitive biological studies.

Purpose of the Study:

  • To present a microwave-assisted protocol for biological sample preparation.
  • To optimize the dehydration and resin embedding steps for TEM.
  • To significantly reduce the overall processing time for biological specimens.

Main Methods:

  • Utilized microwave processors for accelerated sample treatment.
  • Developed a protocol for dehydration of biological tissues.
  • Implemented resin embedding techniques compatible with microwave processing.

Main Results:

  • Achieved complete processing, dehydration, and resin embedding in 4 hours or less.
  • Successfully prepared biological specimens suitable for sectioning and TEM analysis.
  • Demonstrated the efficacy of microwave technology in speeding up sample preparation.

Conclusions:

  • Microwave-assisted processing offers a substantial time advantage for TEM sample preparation.
  • This protocol is effective for a wide range of biological materials.
  • The rapid turnaround time facilitates timely research in electron microscopy.