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Related Experiment Videos

A method for improved resolution for fluorescence microscopy using plastic-embedded material subjected to resin

S D Russell1

  • 1Department of Botany and Microbiology, University of Oklahoma, Norman 73019-0245.

Stain Technology
|January 1, 1990
PubMed
Summary

This study presents a new protocol for extracting epoxy resins from semithin sections using common solvents. This method enhances fluorescence microscopy by minimizing fading and grain, improving image quality.

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Area of Science:

  • Microscopy and Imaging Science
  • Materials Science
  • Chemical Analysis

Background:

  • Semithin sections are crucial for high-resolution microscopy.
  • Traditional methods for preparing sections for fluorescence microscopy can lead to image fading and unacceptable grain.
  • Epoxy resins are commonly used embedding media, but their extraction can be challenging.

Purpose of the Study:

  • To develop an effective protocol for extracting epoxy resins from semithin sections.
  • To improve the quality of fluorescence microscopic observations.
  • To minimize image fading and photographic grain without antifading agents.

Main Methods:

  • Extraction of epoxy resins using sodium hydroxide (NaOH), benzene, acetone, and methanol.
  • Preparation of semithin sections for microscopy.

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  • Utilizing ultrarapid films (e.g., Kodak T-Max P3200) at ISO 3200 for imaging.

    • Main Results:

    • The developed protocol successfully extracts epoxy resins from semithin sections.
    • Semithin sections prepared with this method show superior performance for fluorescence microscopy compared to paraffin or unsectioned materials.
    • Use of high-speed films at ISO 3200 effectively minimizes fading and maintains acceptable photographic grain size.

    Conclusions:

    • The protocol provides a reliable method for preparing samples for enhanced fluorescence microscopy.
    • Semithin sections are advantageous for detailed microscopic analysis.
    • The combination of specific solvents and rapid film photography offers a practical solution for high-quality imaging.