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    PRACTICAL 4: The Staining of Microorganisms - Capsule Staining and Spore Staining

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    Syamim Lazim
    This document discusses techniques for staining bacterial endospores and capsules. It describes how the Schaeffer-Fulton method uses malachite green to stain endospores within vegetative cells, while decolorizing the vegetative cells with water. This allows distinction between endospores and vegetative cells. It also explains how capsule staining uses an acidic stain for the background and a basic stain for bacterial cells, leaving any capsule unstained and visible as a clear halo against the stained background and cells.

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    PRACTICAL 4: The Staining of Microorganisms - Capsule Staining and Spore Staining

    Uploaded by

    Syamim Lazim
    23 views2 pages
    This document discusses techniques for staining bacterial endospores and capsules. It describes how the Schaeffer-Fulton method uses malachite green to stain endospores within vegetative cells, while decolorizing the vegetative cells with water. This allows distinction between endospores and vegetative cells. It also explains how capsule staining uses an acidic stain for the background and a basic stain for bacterial cells, leaving any capsule unstained and visible as a clear halo against the stained background and cells.

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    PRACTICAL 3

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    This document discusses techniques for staining bacterial endospores and capsules. It describes how the Schaeffer-Fulton method uses malachite green to stain endospores within vegetative cells, while decolorizing the vegetative cells with water. This allows distinction between endospores and vegetative cells. It also explains how capsule staining uses an acidic stain for the background and a basic stain for bacterial cells, leaving any capsule unstained and visible as a clear halo against the stained background and cells.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    23 views2 pages

    PRACTICAL 4: The Staining of Microorganisms - Capsule Staining and Spore Staining

    Uploaded by

    Syamim Lazim
    This document discusses techniques for staining bacterial endospores and capsules. It describes how the Schaeffer-Fulton method uses malachite green to stain endospores within vegetative cells, while decolorizing the vegetative cells with water. This allows distinction between endospores and vegetative cells. It also explains how capsule staining uses an acidic stain for the background and a basic stain for bacterial cells, leaving any capsule unstained and visible as a clear halo against the stained background and cells.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 2

    PRACTICAL 4: The Staining of Microorganisms – Capsule Staining and Spore

    Staining

    1.0 INTRODUCTION

    When vegetative cells of certain bacteria such as Bacillus spp and Clostridium spp


    are subjected to environmental stresses such as nutrient deprivation, they produce
    metabolically inactive or dormant form-endospore. Formation of endospore
    circumvent the problems associated with environmental stress and helps them
    to survive. During unfavorable conditions (especially when carbon and nitrogen
    become unavailable) endospores can form within different areas of the vegetative
    cell. They can be central, subterminal, or terminal.  Central endospores are located
    within the middle of the vegetative cell.  Terminal endospores are located at the end
    of the vegetative cell.  Sub-terminal endospores are located between the middle and
    the end of the cell.

    A differential staining technique (the Schaeffer-Fulton method) is used to distinguish


    between the vegetative cells and the endospores.  A primary stain (malachite
    green) is used to stain the endospores.  Because endospores resist staining, the
    malachite green will be forced into (i.e, malachite green permeate the spore wall) the
    endospores by heating.  In this technique heating acts as a mordant. There is no
    need of using any decolorizer in this spore staining as the primary dye malachite
    green bind relatively weakly to the cell wall and spore wall .In fact If washed well with
    water the dye come right out of cell wall however not from spore wall once the dye is
    locked in. Water is used to decolorize the vegetative cells.

     Capsule stain is a type of differential stain which uses acidic and basic dyes to stain
    background & bacterial cells respectively so that presence of capsule is easily
    visualized.  Capsule is synthesized in the cytoplasm and secreted to the outside of
    the cell where it surrounds the bacterium.  Most of the capsulated bacteria have a
    capsule made up of polysaccharide layer but some bacteria have capsule made up
    of polypeptide, or glycoprotein. Bacterial capsules are non-ionic, so neither acidic nor
    basic stains will adhere to their surfaces. Therefore, the best way to visualize them is
    to stain the background using an acidic stain (e.g., Nigrosine, congo red) and to stain
    the cell itself using a basic stain (e.g.,crystal violet, safranin, basic fuchsin and
    methylene blue). Crystal violet and india ink are used. The capsule is seen as a clear
    halo around the microorganism against the black background. This method is used
    for demonstrating Cryptococcus. The background will be dark (color of india ink).The
    bacterial cells will be stained purple (bacterial cells takes crystal violet-basic dyes as
    they are negatively charged). The capsule (if present) will appear clear against the
    dark background (capsule does not take any stain).

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