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Related Experiment Videos

A simple staining method for chromatin in electron microscopy compatible with serial sectioning.

C Esquivel, P Rovira, O Echeverría

    Ultramicroscopy
    |January 1, 1987
    PubMed
    Summary
    This summary is machine-generated.

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    This study presents a modified phosphotungstic acid (PTA) staining method for improved chromatin visualization in electron microscopy. The new protocol enables epoxy resin embedding, facilitating detailed nuclear structure analysis.

    Area of Science:

    • Cell Biology
    • Microscopy Techniques
    • Molecular Biology

    Background:

    • Studying interphasic nucleus chromatin disposition requires specific staining and serial sectioning.
    • Traditional phosphotungstic acid (PTA) staining was difficult with glycolmethacrylate resins, hindering serial sectioning.
    • Alternative embedding methods are needed for effective chromatin analysis.

    Purpose of the Study:

    • To develop an improved phosphotungstic acid (PTA) staining protocol for chromatin.
    • To enable the use of epoxy resins for electron microscopy of nuclear chromatin.
    • To enhance the study of chromatin disposition within the interphasic nucleus.

    Main Methods:

    • Tissue fixation using 2.5% glutaraldehyde at pH 7.2.
    • Acidic rinsing (0.2N HCl, pH 2.1-2.3) and staining with 3% PTA in 1N HCl.

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  • Dehydration in ethanol and embedding in Epon resin.

    • Main Results:

    • A modified PTA staining procedure was established.
    • The method allows for successful epoxy resin embedding of tissues.
    • This technique is compatible with serial sectioning for electron microscopy.

    Conclusions:

    • The developed protocol enhances chromatin staining for electron microscopy.
    • This method overcomes limitations of previous techniques, improving nuclear structure studies.
    • The improved staining facilitates detailed analysis of chromatin organization.