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    Home > Biochemistry News > Microbiology News > Bacterial membrane staining

    Bacterial membrane staining

    • Last Update: 2021-01-21
    • Source: Internet
    • Author: User
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    (i) Experimental purpose
    : learning the bacteria's membrane staining method:
    (ii) experimental principle:
    because of the affinity between the diaphragm and dye, not easy to color, usually using negative staining method to dye the membrane, that is, try to make the bacteria and background coloring and membrane does not color, so that the membrane around the bacteria body is a transparent circle. Because the water content of the film is more than 90%, so dyeing is generally not
    heating
    fixed, so as to avoid the film wrinkle deformation.
    (iii) the test equipment
    1. Living materials:
    culture
    glial bud bacillus (Bacillus mucilaginosus, commonly known as "potassium bacteria") for 3-5 days. When the bacteria grow on the carbon-based
    of glycol
    , the membrane is abundant.
    2. Staining fluid and
    reagents
    :Tyler staining fluid: (see annex II, (i), 14), drawing ink after
    filtration paper
    filtration
    and red staining fluid (see Annex II, (i), 2), melanin (see Annex II, (i), 7), 6% glucose aqueous solution, 1% methyl purple aqueous solution, methanol, 20% CuSO4 aqueous solution, balsamic asphalt, xylene.
    3. Equipment: slides, slide shelves, wipe paper,
    microscopes,
    etc.
    (iv) Experimental method
    recommends the following four dyeing methods, of which the wet ink method is simpler and suitable for a variety of bacteria with pods. This is even more effective if examined with a difference microscope.
    1. Negative staining method:
    (1) production: take a piece of clean slide, add a drop of distilled water, take a small amount of bacteria into the water droplets mixed and coated.
    (2)
    dry
    : dry the smears in the air or dry them with a dry air blow.
    (3) staining: add red staining liquid staining 2-3min on the coating surface.
    (4) wash: wash the red dye with water.
    (5) dry: put the staining sheet in the air to dry or dry with a cold air blow.
    (6) Melanin: Add a small drop of melanin to the left side of the dye coating surface, gently touch the melanin with an edge smooth slide, spread the melanin along the edge of the slide, and then drag to the right to make the melanin a thin layer on the dye coating surface and quickly dry.
    (7) mirror inspection: first low mirror, then high mirror observation.
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