Special Staining; spore and capsule PowerPoint Presentation, PPT - DocSlides

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Special Staining; spore and capsule PowerPoint Presentation, PPT - DocSlides - Lab #4 . Medgar Evers College. Bio 261 Microbiology. Prof. Santos. Aim #1 exercise 14. Capsular staining. Capsule- extracellular layer composed of polysaccharides found in some bacteria that plays a role in immunity and adhesion..

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Special Staining; spore and capsule PowerPoint Presentation, PPT - DocSlides

    Slide1

    Special Staining; spore and capsule

    Lab #4

    Medgar Evers College

    Bio 261 Microbiology

    Prof. Santos

    Slide2

    Slide3

    Aim #1 exercise 14

    Capsular staining

    Capsule- extracellular layer composed of polysaccharides found in some bacteria that plays a role in immunity and adhesion.

    It helps bacteria like

    Streptococcus pneumonia

    by preventing white blood cells from engulfing them and allowing them to invade the lungs.

    It helps bacteria like

    Streptococcus mutans

    attach to the surface of teeth and cause dental plaque.

    Slide4

    Problems staining the capsule

    1- fixing the cells after creating a smear will destroy the delicate capsule.

    2- If cells are not fixed properly, they will wash off during staining, thus you will not see anything!

    We avoid this by using a combined method of negative staining and a simple stain.

    Slide5

    procedure

    1- we mix a couple of loopfuls of the

    Klebsiella pneumonia

    with India Ink. This will stain the background.

    2- The ink suspension is spread over the slide and air dried.

    3- GENTLY heat fix!

    4- stain with crystal violet for one minute!

    5- wash the crystal violet off with water

    6- blot dry and look under microscope!

    Slide6

    Slide7

    Aim #2 Exercise 16

    Spore stain

    Some species of bacteria belonging to the genera

    Bacillus

    and

    Clostridia

    produce tough protective structures called endospores.

    Endospores are resting stages that allow the bacteria to resist harsh environmental conditions or treatments.

    The spore contains a tough protein coat called the exosporium that allows it to form a barrier against all harsh conditions.

    A spore is a very dehydrated structure (as much as 10 to 30 % less water than vegetative cell). The less water it has the more heat resistant.

    Slide8

    Due to nature of the spore, it is hard to stain them and get the stain to penetrate. But, using heat as the mordant allows the primary stain to penetrate the spore.

    2 methods; Schaeffer-Fulton method and Dorner method

    Slide9

    Schaeffer- Fulton

    Malachite green is the primary stain and stains the endospore greenSafranin is the counterstain and stains the vegetative cell pink.

    Slide10

    Dorner method

    Only the spore is stained red with carbolfuchsin and the background is treated with Nigrosin to create contrast.

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