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Transmission Electron Microscopy01:15

Transmission Electron Microscopy

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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

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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

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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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Preparation of Samples for Electron Microscopy01:20

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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 Microscopy of Bone.

Vincent Everts1,2, Anneke Niehof3,4, Wikky Tigchelaar-Gutter5

  • 1Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam, VU University Amsterdam, Amsterdam, The Netherlands. v.everts@acta.nl.

Methods in Molecular Biology (Clifton, N.J.)
|February 8, 2019
PubMed
Summary
This summary is machine-generated.

Transmission electron microscopy (TEM) is crucial for analyzing mineralized tissues such as bone and dentin. This chapter details methods for preparing these tissues for ultrastructural examination, aiding in understanding tissue organization and interactions.

Keywords:
BoneMineralized tissueTransmission electron microscopyUltrastructure

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Area of Science:

  • Biomineralization research
  • Cellular and tissue ultrastructure
  • Materials science of biological tissues

Background:

  • Understanding the ultrastructure of mineralized tissues like bone and dentin is vital.
  • Cell-cell and cell-matrix interactions within these tissues require detailed analysis.
  • Three-dimensional organization impacts tissue function and integrity.

Purpose of the Study:

  • To describe methods for preparing mineralized tissues for electron microscopy.
  • To facilitate ultrastructural analysis of bone and dentin.
  • To enhance the study of interactions and organization in mineralized tissues.

Main Methods:

  • Sample preparation techniques for mineralized tissues.
  • Processing for transmission electron microscopy (TEM).
  • Adaptation of methods for diverse tissue sources.

Main Results:

  • Established protocols for mineralized tissue processing.
  • Demonstrated feasibility of TEM for ultrastructural details.
  • Enabled visualization of cell-cell/cell-matrix interactions.

Conclusions:

  • Effective methods exist for preparing mineralized tissues for TEM.
  • Ultrastructural analysis provides key insights into tissue organization.
  • These techniques are essential for advancing biomineralization research.