Endospore and Capsule Staining Methods

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Questions and Answers

What is the purpose of applying Malachite Green in the Schaeffer-Fulton Method?

  • To prepare the bacterial culture
  • To visualize endospores (correct)
  • To dry the slide quickly
  • To stain vegetative cells only

Endospores stain red when using the Schaeffer-Fulton Method.

False (B)

What is the first step in capsule staining using Maneval’s Method?

Prepare the smear with India ink or Congo red.

In the endospore staining process, the slide should air dry after __________ and before counterstaining with Safranin.

<p>rinsing with water</p> Signup and view all the answers

Match each stain to its purpose in the given staining methods:

<p>Malachite Green = Staining endospores Safranin = Counterstaining vegetative cells Maneval’s Stain = Staining bacterial cells in capsules India Ink = Providing a dark background</p> Signup and view all the answers

How long should the Malachite Green stain sit on the smear?

<p>5-10 minutes (B)</p> Signup and view all the answers

Capsules will appear red against a dark background when using the Maneval’s Method.

<p>False (B)</p> Signup and view all the answers

What is used to create a dark background in the capsule staining method?

<p>India ink or Congo red</p> Signup and view all the answers

After rinsing the slide with water during endospore staining, it should be __________ before examining under a microscope.

<p>blotted dry</p> Signup and view all the answers

Which lens is used to examine the slides in the staining methods described?

<p>Oil immersion lens (C)</p> Signup and view all the answers

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Study Notes

Endospore Staining (Schaeffer-Fulton Method)

  • Begin with a bacterial smear on a slide, allowing it to air dry before heat-fixing by passing through a flame multiple times.
  • Position the slide over a steaming water bath and cover the smear with a small piece of paper towel.
  • Flood the smear with malachite green stain and allow it to sit for 5-10 minutes, keeping the stain moist by adding more if necessary.
  • After steaming, cool the slide and gently rinse with water to remove excess malachite green.
  • Apply safranin as a counterstain, letting it sit for 30 seconds to 1 minute.
  • Rinse the slide again to remove excess safranin and blot it dry using absorbent paper.
  • Under an oil immersion lens, endospores will appear green while vegetative cells will appear red.

Capsule Staining (Maneval’s Method)

  • Start by placing a drop of India ink or Congo red on one end of a clean slide.
  • Mix a small amount of bacterial culture into the drop and spread it thinly using another slide's edge; allow it to air dry without heat-fixing.
  • Once dry, flood the smear with Maneval’s solution and let it sit for 1-2 minutes.
  • Gently rinse with water to eliminate excess Maneval’s stain, avoiding disruption of the smear.
  • Allow the slide to air dry completely without blotting or heat application.
  • Under an oil immersion lens, capsules will be visible as clear halos around the pink/red-stained bacterial cells, with a dark background from the India ink or Congo red.

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