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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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Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
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Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
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Related Experiment Video

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Bacterial Immobilization for Imaging by Atomic Force Microscopy
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A cost-effective method to immobilize hydrated soft-tissue samples for atomic force microscopy.

Suchit Sahai1, Marysuna Wilkerson1, Ana Maria Zaske2

  • 1Department of Pediatric Surgery, McGovern Medical School, UTHealth - The University of Texas Health Science Center at Houston, Houston, TX.

Biotechniques
|October 8, 2016
PubMed
Summary

Researchers developed a cost-effective method using transglutaminase to immobilize hydrated soft tissue specimens for atomic force microscopy (AFM). This technique is significantly cheaper than existing methods, offering a practical solution for AFM tissue characterization.

Keywords:
Wharton’s jellyatomic force microscopybioadhesivespolyphenolic proteinssoft tissuetransglutaminasewet-adhesion

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Area of Science:

  • Biomaterials Science
  • Biophysics
  • Tissue Engineering

Background:

  • Immobilizing hydrated soft tissue specimens for atomic force microscopy (AFM) presents a significant challenge.
  • Existing methods may be costly or less effective for maintaining tissue hydration during analysis.

Purpose of the Study:

  • To develop and validate a simple, cost-effective method for immobilizing hydrated soft tissue specimens for AFM.
  • To compare the efficacy and cost of transglutaminase-based immobilization with a commercial tissue adhesive.

Main Methods:

  • Utilized transglutaminase in an aqueous environment for tissue immobilization.
  • Applied the method to human native Wharton's Jelly (nWJ) specimens.
  • Performed side-by-side comparison with Corning Cell-Tak, a commercial polyphenolic protein-based adhesive.

Main Results:

  • Transglutaminase successfully immobilized hydrated nWJ specimens for AFM characterization.
  • Both transglutaminase and Cell-Tak effectively immobilized nWJ in its physiological hydrated state.
  • Transglutaminase demonstrated a cost that is over 3000-fold lower than Cell-Tak.

Conclusions:

  • Transglutaminase offers a highly cost-effective and efficient method for immobilizing hydrated soft tissues for AFM.
  • This approach provides an attractive alternative for researchers requiring reliable tissue immobilization for AFM analysis.
  • The method preserves the native hydrated state of the tissue, crucial for accurate AFM characterization.