(LAB) Negative Stain Technique of Bacterial Capsules
Terms in this set (24)
Does all bacteria have capsules?
How are capsules anti-phagocytic
They limit ability of phagocyte to engulf bacteria
How else do capsules benefit bacteria?
Help with adherence to host tissues (cells)
Differentiating between bacteria which leads to proper disease diagnosis
What is the negative stain technique also called?
What structure of bacteria are you "indirectly" looking for?
How do you know if a capsule is really present?
Will see a "halo" around the purple bacterium
What's the color of the background in negative/capsule staining?
Why doesn't the halo pick up dye/not stained?
It has no charge because polysaccharide is uncharged; can't pick-up charged dyes = won't stain
What stains are used in this technique?
India ink and crystal violet
Gently resuspend culture
What does india ink stain?
Background; is a non-ionic dye
Put 1 drop of india ink at one end of slide
Put 2 loops of nutrient broth culture next to ink drop (flame between loops)
Mix all 3 drops at the end of slide
Take 2nd slide (pushing slide) at an angle on the mixture of 1 st slide => push and smear evenly along 1st slide
Air dry 1st slide (DON'T HEAT FIX)
What does crystal violet stain?
Bacterium inside the capsule
Cover entire slide with crystal violet for 60 seconds
Rinse slide with water
Blot and air dry slide
Observe under oil immersion lens
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