Endospore staining by Bartholomew and Mittwer’s Method
To stain Endospore of a bacterial cell by Bartholomew and Mittwer’s Method.
- Cell suspension of endospore producing bacteria.
- Malachite green stain.
- Saffranin stain.
- Take a clean grease free slide and prepare a thick smear on a slide.
- The smear is heat fixed by passing the slide from the flame for about 25 times.
- The slide is allowed to cool.
- Further the slide is treated with Malachite green stain and allowed it to react for about 10 minutes.
- After 10 minutes slide is given a water wash treatment.
- Further the slide is treated with counter stain that is saffranin for about 30 seconds.
- After 30 seconds the slide is water washed, air dried and observed under oil immersion.