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  • Gram and ZN Stain PDF

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    The document provides instructions for Ziehl Neelsen staining and Gram staining procedures. For Ziehl Neelsen staining, slides are fixed, stained with carbol fuchsin, decolorized with acid, and counterstained to identify acid-fast bacilli as pink rods. For Gram staining, slides are fixed, stained with crystal violet, iodine, acetone, and safranin to distinguish Gram-positive bacteria as violet and Gram-negative as red under a microscope.

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    The document provides instructions for Ziehl Neelsen staining and Gram staining procedures. For Ziehl Neelsen staining, slides are fixed, stained with carbol fuchsin, decolorized with acid, and counterstained to identify acid-fast bacilli as pink rods. For Gram staining, slides are fixed, stained with crystal violet, iodine, acetone, and safranin to distinguish Gram-positive bacteria as violet and Gram-negative as red under a microscope.

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    © All Rights Reserved
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    28 views1 page

    Gram and ZN Stain PDF

    Uploaded by

    waheeda
    The document provides instructions for Ziehl Neelsen staining and Gram staining procedures. For Ziehl Neelsen staining, slides are fixed, stained with carbol fuchsin, decolorized with acid, and counterstained to identify acid-fast bacilli as pink rods. For Gram staining, slides are fixed, stained with crystal violet, iodine, acetone, and safranin to distinguish Gram-positive bacteria as violet and Gram-negative as red under a microscope.

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    of 1

    ZIEHL NEELSEN STAINING

    1. Make smear and fix it by passing the slide rapidly over the flame. Keep the slide
    horizontal (not tilting).

    2. Apply pure carbol fuchsin (not safranine) for 5 minutes .

    3. Heat up the slide. Boiling must be avoided. Wash with water.

    4. Apply 20% H2SO4 (suphuric acid) for 4 minute (till no redness).

    5. Wash with water.

    6. Apply methylene blue (or Malachite green) for 15-20 Sec.

    7. Wash with water.

    8. Dry in air and see under microscope using oil immersion – Lens 100.

    RESULT:- TO SEE AFB as Pink rods Lt Col Nasrullah Malik

    GRAM STAINING
    1. Make smear and fix it by passing the slide over the flame. Keep the slide horizontal
    (not tilting).

    2. Apply crystal violet for 30 sec.

    3. Wash with water.

    4. Apply gram iodine for 30 sec.

    5. Wash with water.

    6. Apply Acetone iodine for 30 sec.

    7. Wash with water.

    8. Apply dilute 1/10 carbol fuchsin for 30 sec.


    9. Wash with water.
    10. Dry and see under microscope. Use oil immersion lens.

    RESULT:- Gram + = Violet Gram Neg = Red LT. COL. Nasrullah

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