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Artifacts associated with quick-freezing and freeze-drying.

K R Miller, C S Prescott, T L Jacobs

    Journal of Ultrastructure Research
    |February 1, 1983
    PubMed
    Summary
    This summary is machine-generated.

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    Quick-freezing and freeze-drying can create artifactual filament structures in biological samples. Researchers must use caution and wash samples in distilled water to accurately study cell architecture.

    Area of Science:

    • Cryo-electron microscopy
    • Biophysics
    • Materials Science

    Background:

    • Quick-freezing and freeze-drying are common techniques for preparing biological samples for microscopy.
    • Artifact formation during sample preparation can lead to misinterpretation of cellular structures.

    Purpose of the Study:

    • To investigate the structures formed by quick-freezing and freeze-drying in nonbiological and biological samples.
    • To assess the potential for artifact generation during this sample preparation method.

    Main Methods:

    • Utilized a liquid helium-cooled freeze-slamming device for rapid freezing.
    • Examined nonbiological samples (sodium chloride, sucrose, Tris buffer) and biological samples (isolated chloroplast membranes).
    • Prepared samples in distilled water and various salt solutions.

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    Main Results:

    • Nonbiological samples formed filament-like and trabeculum-like structures, possibly eutectic mixtures.
    • Biological membranes in salt solutions appeared embedded in a filament network resembling a cytoskeleton.
    • Excellent membrane surface replicas were obtained in distilled water.

    Conclusions:

    • The quick-freezing and freeze-drying technique can introduce artifacts, particularly in salt-containing buffers.
    • Extreme caution is advised when interpreting cell architecture prepared with this method.
    • Washing biological components in distilled water may be necessary for accurate structural representation.