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Preservation and contrast without osmication or section staining

M Locke1

  • 1Department of Zoology, University of Western Ontario, London, Canada.

Microscopy Research and Technique
|September 1, 1994
PubMed
Summary
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Uranyl acetate (UA) offers a superior alternative to osmium tetroxide for electron microscopy tissue preparation. Aldehyde fixation followed by UA preserves cellular structures and chemical activity for advanced labeling techniques.

Area of Science:

  • Electron Microscopy
  • Cell Biology
  • Biochemistry

Background:

  • Conventional electron microscopy uses aldehyde fixation and osmium tetroxide (OsO4) postfixation.
  • OsO4 postfixation, while yielding aesthetically pleasing results, can degrade cellular components and reduce chemical reactivity, impacting downstream applications like antibody and lectin binding.

Purpose of the Study:

  • To evaluate an alternative tissue preparation method for transmission electron microscopy.
  • To assess the preservation of cellular structures and chemical activity using uranyl acetate (UA) as a postfixative agent.

Main Methods:

  • Tissues were fixed with aldehydes.
  • Postfixation was performed using uranyl acetate (UA) instead of osmium tetroxide.
  • Microscopy and chemical reactivity assays (immunogold and lectin-gold labeling) were conducted.

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Main Results:

  • Aldehyde/UA fixation preserved cellular ultrastructure effectively.
  • This method enhanced contrast, allowing observation without section staining.
  • Chemical activity, crucial for immunogold and lectin-gold labeling, was retained.

Conclusions:

  • Aldehyde fixation followed by UA postfixation is a viable alternative to conventional OsO4 treatment.
  • This method improves preservation and contrast while maintaining chemical reactivity for advanced labeling in electron microscopy.