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    Preservation method for long-term storage of fluorescently labeled cells for microscopy

    Jacob Robertson, Erin Garza

    Published: 2024-01-12 DOI: 10.17504/protocols.io.ewov1q15pgr2/v1
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    Abstract

    This method can be used to preserve fluorescently labeled bacterial cells for long-term storage before imaging on a fluorescent microscope. Using this protocol, samples can be saved at 4°C for weeks-months while maintaining strong, easily detectable fluorescence and cell integrity.

    Before start

    Prepare the preservation components: 10% glycerol (sterile), 4% paraformaldehyde in PBS pH 7.4 (sterile)

    Steps

    Sample

    1.

    Collect your cells (up to 500 μl) in a sterile microfuge tube.

    • This protocol can also be used on solid material containing biofilms.

    Preserve

    2.

    To your sample tube, add 250µL sterile 10% glycerol and 250µL sterile 4% paraformaldehyde in PBS (pH 7.4).

    • If preserving solid material, make sure the entire sample is submerged in the preservation solution.
    3.

    Gently Mix

    Storage

    4.

    Store samples at 4°C until ready to image.

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