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Scanning Electron Microscopy01:07

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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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Visualizing cell-laden fibrin-based hydrogels using cryogenic scanning electron microscopy and confocal microscopy.

Maya Schnabel-Lubovsky1,2, Olga Kossover1, Sonia Melino3

  • 1Department of Biomedical Engineering, Technion-Israel Institute of Technology, Haifa, Israel.

Journal of Tissue Engineering and Regenerative Medicine
|January 20, 2019
PubMed
Summary

This study introduces a new imaging method for high-water-content hydrogels. Cryogenic high-resolution scanning electron microscopy (HR-SEM) visualizes cells and biomaterials in their natural state, improving tissue engineering strategies.

Keywords:
confocal microscopyelectron microscopyfibrinhydrogelscaffoldtissue engineering

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Area of Science:

  • Biomaterials Science
  • Cell Biology
  • Microscopy

Background:

  • Cell-laden hydrogels with high water content (>98%) are crucial for tissue engineering.
  • Imaging these delicate, hydrated structures presents significant challenges due to potential artifacts.

Purpose of the Study:

  • To develop and validate an imaging strategy for high-resolution visualization of cell-laden hydrogels.
  • To minimize imaging artifacts and preserve the native hydrated state of the hydrogel constructs.

Main Methods:

  • Confocal microscopy was used for initial imaging.
  • Cryogenic high-resolution scanning electron microscopy (HR-SEM) was employed to achieve submicron and subcellular resolution.
  • Imaging was performed on poly(ethylene glycol)-fibrinogen and fibrin hydrogels with embedded human dermal fibroblasts.

Main Results:

  • Cryogenic HR-SEM successfully visualized ultrastructural details, including cellular components and the cell-biomaterial interface.
  • The native hydrated state of the hydrogel and the integrity of cell membranes were preserved.
  • Interactions between cells, albumin microbubbles, and the hydrogel matrix were detailed.

Conclusions:

  • The developed imaging strategy provides a comprehensive view of cell-laden hydrogels in their hydrated state.
  • This methodology overcomes challenges in imaging high-water-content hydrogels, offering valuable insights for tissue engineering and regeneration.