21.4: Materials and Procedures

Bio 221Lab: Introduction to Microbiology · LibreTexts

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    From LibreTexts, under CC BY, as the source states it; its equations are written in TeX, which the PDF shows typeset. This page at LibreTexts.

    Materials

    • Student Stock Cultures:
      • Mycobacterium smegmatis
      • Staphylococcus saprophyticus
    • Glass Slides
    • Stain tray and slide holder
    • Stain kit:
      • Kinyoun carbolfuchsin
      • Acid/Alcohol decolorizer
      • Methylene blue
      • Glass bowls with Sanisol

    Procedures

    1. Prepare a heat-fixed smear prep with a mixture of Mycobacterium smegmatis and Staphylococcus saprophyticus. Aseptically place a drop of each onto the same spot on the slide and mix well. Try to break up the waxy Mycobacterium smegmatis so that it can spread evenly across the slide. Air dry, then heat-fix.
    2. Cover the smear with Kinyoun carbolfuchsin and let stand for 5-10 minutes.
    3. Gently wash with DI water.
    4. Gently decolorize with Acid-Alcohol by dropping the decolorizer down the slide until the slide runs clear. Note that this decolorizer is not the same as the Acetone-Alcohol used in the Gram stain.
    5. Gently wash with DI water.
    6. Counterstain by covering the slide with Methylene blue for 1 min.
    7. Gently wash with DI water, blot, and view under oil immersion.
    Instructions for applying silicone adhesive, detailing steps with illustrations for clarity.
    Figure \(\PageIndex{1}\): Slide Prep, C.S.Ramey, CC-BY-NC-SA

    Results

    1. Draw and color (or indicate colors of) your Acid-Fast stain:
    Blank diagram for organizing information with a circular area and fields for organizer details, size, and magnification.

    This page titled 21.4: Materials and Procedures was last modified on Sat, 16 May 2026 01:44:52 GMT and is shared under a CC BY license and was authored, remixed, and/or curated by Kelly C. Burke.