Endospore staining by Bartholomew and Mittwer’s Method

Endospore staining by Bartholomew and Mittwer’s Method

Aim

To stain Endospore of a bacterial cell by Bartholomew and Mittwer’s Method.

Requirement

  1. Cell suspension of endospore producing bacteria.
  2. Malachite green stain.
  3. Saffranin stain.

Procedure

  1. Take a clean grease free slide and prepare a thick smear on a slide.
  2. The smear is heat fixed by passing the slide from the flame for about 25 times.
  3. The slide is allowed to cool.
  4. Further the slide is treated with Malachite green stain and allowed it to react for about 10 minutes.
  5. After 10 minutes slide is given a water wash treatment.
  6. Further the slide is treated with counter stain that is saffranin for about 30 seconds.
  7. After 30 seconds the slide is water washed, air dried and observed under oil immersion.

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