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3.10: Acid-Fast Stain

  • Page ID
    90987
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    Preparation

    1. Have test tubes with sterile TSB on hand.
    2. 12-18 hours before the laboratory, inoculate test tubes of sterile TSB with a mixture of Staphylococcus epidermidis and Mycobacterium chelonae and incubate at 37 °C .

    Materials

    • 12-18 hour cultures (enough for groups to share)
    • plain microscope slides (at least 1 per group)
    • Bunsen burners (1 per group)
    • inoculating loops (1 per group)
    • (if using bacteria on solid medium rather than TSB) droppers with saline, DI water, or distilled water
    • slide warmer
    • test tube racks (1 per group plus one or more racks to hold cultures)
    • 400-500 mL beakers with ~200 mL of water each (1 per group)
    • hot plate (1 per group)
    • slide rack hat fits over beaker (1 per group)
    • paper towels
    • carbol fuchsin in dropper bottles (1 per group)
    • acid-alcohol in dropper bottles (not ethanol from the Gram stain kit) (1 per group)
    • methylene blue in dropper bottles (1 per group)
    • wood test tube clamps/holders (1 per group)
    • small chemical waste containers (1 per group)
    • distilled or DI water in squirt bottles (1 per group)
    • bibulous paper (enough for each groups to blot their slides)
    • light microscopes (1 per group)
    • colored pencils
    • (optional, but recommended) Wax pencils (1 per group)
    • (if using oil immersion) immersion oil dropper bottles
    • (if using oil immersion) lens paper

    This page titled 3.10: Acid-Fast Stain is shared under a CC BY-NC-SA 4.0 license and was authored, remixed, and/or curated by Rosanna Hartline.

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