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    Smearing and Gram

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    Basma Mohamed

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    © All Rights Reserved
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    Smearing and Gram

    Uploaded by

    Basma Mohamed
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    © © All Rights Reserved

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    Copyright:

    © All Rights Reserved
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    Download as pptx, pdf, or txt
    8 views7 pages

    Smearing and Gram

    Uploaded by

    Basma Mohamed

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    You are on page 1of 7

    Preparation of Bacterial Smears

    and Gram Staining


    The Department of Microbiology
    Faculty of Vet. Medicine
    Cairo University
    April, 2021
    How to make a bacterial smear
    1. Label a clean glass slide as demonstrated by your instructor.
    2. Add a small drop of water to the slide using the inoculating loop.
    3. Pick up a small amount of bacteria.
    4. Mix it well with the water and spread the mixture over a wider area of
    the slide.
    5. Air dry the bacterial specimen on the slide.
    6. Heat fix the bacterial specimen by passing the slide slowly over the
    flame twice.
     Heat fixing kills cells, and adheres them to the slide.
     Cells will be rinsed off the slides if they are not heat fixed properly.
     Be careful not to overheat the slides in this procedure

    After heat-fixing is complete, you are ready to stain your slide.


    Gram’s staining method
    1)Flood the sample with crystal violet solution and let it stand for 30- 90 seconds.

    2)Gently wash the stain off with running tap water for~5 seconds. Shake off excess
    water.

    3)Flood the smear with Gram’s iodine solution for 1min.

    4)Gently wash the stain off with running tap water for~5 seconds. Shake off excess
    water.

    5)Decolourize with decolourizing solution by slowly dropping the solution above the
    smear on the slightly tilted slide.

    6)Gently rinse off the stain with running water for ~5 seconds. Shake off excess water.

    7)Cover the slide with counterstain for 30 seconds.

    8)Gently rinse off the stain with running water for ~5seconds. Shake off excess water.

    9) Carefully blot with filter paper, put a drop of cidar wood oil and examine with oil
    immersion lens.
    If you're sure that you have mixed preparation of G+ and G-
    then you should see purple and pink cells

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