Explain Procedure for Spore Staining in a Given Bacterial Culture

Carry out the exercise following the steps enumerated herewith

1. Take a clean, non-greasy slide. Prepare bacterial smear by taking a small amount of culture and mixing it with a drop of water on a slide. Spread it in a rounded manner.

2. Air dry and heat fix the smear.

3. Keep the slide on a beaker having boiling water. Flood the smear with malachite green and allow the steam to pass through it for 4-5 minutes. Replenish the stain if it starts evaporating and drying.

4. Remove the slide from the beaker and allow it to cool.

5. Decolourize the smear by washing slowly under running tap water.

6. Counter stain the smear with safranin for 30 seconds.

7. Wash the slide again with tap water.

8. Blot dry the slide with blotting paper and observe under oil immersion.