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  • Negative Staining

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    Özge Kavaklı
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    This document describes the process of negative staining using nigrosin or India ink to view bacterial morphology. The stain does not color the negatively charged bacterial cells, allowing their shapes and sizes to be seen as colorless outlines against the dark background of the stained slide glass. An advantage is that prior heat fixation is not required, so bacteria appear in more natural shapes. The process involves mixing a bacterial sample with the acidic stain, which repels from the negatively charged cells, then smearing and air drying the mixture on a microscope slide for observation.

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    description of negative staining

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    NEGATIVE STAINING

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    This document describes the process of negative staining using nigrosin or India ink to view bacterial morphology. The stain does not color the negatively charged bacterial cells, allowing their shapes and sizes to be seen as colorless outlines against the dark background of the stained slide glass. An advantage is that prior heat fixation is not required, so bacteria appear in more natural shapes. The process involves mixing a bacterial sample with the acidic stain, which repels from the negatively charged cells, then smearing and air drying the mixture on a microscope slide for observation.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Flag for inappropriate content
    10 views1 page

    Negative Staining

    Uploaded by

    Özge Kavaklı
    This document describes the process of negative staining using nigrosin or India ink to view bacterial morphology. The stain does not color the negatively charged bacterial cells, allowing their shapes and sizes to be seen as colorless outlines against the dark background of the stained slide glass. An advantage is that prior heat fixation is not required, so bacteria appear in more natural shapes. The process involves mixing a bacterial sample with the acidic stain, which repels from the negatively charged cells, then smearing and air drying the mixture on a microscope slide for observation.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Flag for inappropriate content
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    NEGATIVE STAINING (SIMPLE-INDIRECT STAINING)

    Nigrosin or India ink is a simple and indirect stain used for determining bacterial morphology.
    The shapes and sizes of the organisms are seen as color-free outlines against the dark
    background.
    An advantage of using this method is that prior fixation by heat is not needed, so the
    organisms are seen in more lifelike shapes.
    Nigrosin or India ink is an acidic stain which becomes negatively charged. Since the surface of
    most bacterial cells is negatively charged, the cell surface repels the stain.
    The glass of the slide will stain, but the bacterial cells will not.

    - Transfer a drop of nigrosin/india ink with a pasteur pipette on the microscope slide (at
    one end of the slide)
    - Using sterilized inoculation loop take a small amount of bacteria (inoculum) and mix
    with stain
    - Using second microscobe slide gently spread the mix (place it near the sample-dye
    mixture at an angle of about 30°- 45°) across the slide. Generate a smear.
    - Allow to air dry
    - Observe under 100x objective

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