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Related Experiment Videos

A high-yield technique for preparing cells fixed in suspension for scanning electron microscopy.

S K Sanders, E L Alexander, R C Braylan

    The Journal of Cell Biology
    |November 1, 1975
    PubMed
    Summary

    This study presents a high-yield method for preparing human leukocytes for scanning electron microscopy (SEM). The technique ensures nearly 100% cell retention, preserving surface architecture for accurate morphological analysis.

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    Area of Science:

    • Cell Biology
    • Microscopy Techniques
    • Biomedical Research

    Background:

    • Scanning electron microscopy (SEM) is crucial for visualizing cell surface architecture.
    • Traditional methods for preparing cells for SEM can lead to cell loss and surface changes.
    • A need exists for a reproducible, high-yield method for processing cells in suspension for SEM.

    Purpose of the Study:

    • To develop and validate a simple, high-yield technique for preparing human leukocytes fixed in suspension for SEM.
    • To prevent alterations in cell surface architecture during sample preparation.
    • To enable quantitative correlations between SEM imaging and other cellular parameters.

    Main Methods:

    • Human leukocytes were fixed in suspension and settled onto poly-L-lysine-coated coverslips.

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  • Cells underwent dehydration in ethanol and critical point drying with CO2.
  • Samples were coated with gold-palladium and observed using SEM, with light microscopy used for cell counting.

    • Main Results:

    • The developed technique achieved a cell yield approaching 100%.
    • No significant changes in surface architecture were observed compared to live cell collection methods.
    • The method demonstrated reproducibility and high efficiency in retaining cells throughout processing.

    Conclusions:

    • This technique offers a simple, reproducible, and high-yield approach for processing fixed cells in suspension for SEM.
    • It effectively preserves cell surface architecture and minimizes selective cell loss.
    • The method facilitates quantitative analysis and correlation with other cellular morphology and function studies.