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Handling and staining epoxy resin sections for light microscopy.

I M Roberts, A M Hutcheson

    Journal of Microscopy
    |January 1, 1975
    PubMed
    Summary
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    This study presents a new method for collecting and staining ultrathin epoxy resin sections for electron microscopy. The technique ensures accurate serial section collection and provides a versatile staining combination for various specimens, even after long-term storage.

    Area of Science:

    • Electron Microscopy Techniques
    • Materials Science
    • Histology

    Background:

    • Standard methods for preparing ultrathin sections for electron microscopy can be challenging.
    • Collecting serial sections without interruption is crucial for detailed ultrastructural analysis.
    • Staining protocols need to be robust and applicable across different specimen types and embedding resins.

    Purpose of the Study:

    • To develop an improved technique for collecting serial ultrathin epoxy resin sections.
    • To establish a reliable and versatile bulk staining method for electron microscopy specimens.
    • To ensure the long-term stability and applicability of the staining protocol.

    Main Methods:

    • Ultrathin epoxy resin sections (0.1-1.0 µm) were collected using cover slip strips from an ultramicrotome.

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  • Sections were dried on a hot plate and bulk stained in custom troughs.
  • A two-step staining process involved periodic acid treatment, followed by basic fuchsin and alkaline methylene blue.

    • Main Results:

    • The handling technique facilitated accurate collection of serial sections without disrupting the sectioning process.
    • The stain combination proved effective for diverse specimens embedded in various epoxy resins.
    • Stained sections remained unaffected by storage for over 1 year, demonstrating excellent stability.

    Conclusions:

    • The developed technique offers a significant improvement for serial section collection in electron microscopy.
    • The proposed bulk staining method is versatile, reliable, and suitable for long-term storage.
    • This approach enhances the efficiency and reproducibility of electron microscopy sample preparation.