20.4: Materials and Procedures
- Page ID
- 40284
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- Student Stock Cultures and saline spore solutions:
- Bacillus cereus
- Bacillus subtilis
- Sporosarcina ureae
- Glass Slides
- Stain tray and slide holder
- Stain kit:
- Malachite Green
- Safranin
- Filter paper rounds
- Hot plate set-up: small beakers of DI water with boiling stones
- Glass bowls with Sanisol
Procedures
- Prepare heat-fixed slides of each culture.
- Place a filter paper round on the slide on top of the smear.
- Cover the filter paper and slide with Malachite Green and at the hot plate set-ups, place the slide across the top of a steaming beaker of DI water. Steam for 5-10 minutes. While steaming, add additional stain as needed to keep the filter paper wet. DO NOT let the slide dry out.
- Remove and discard the filter paper. DO NOT discard in the sink!
- Gently wash with DI water (this is essentially the decolorizer step; there is no separate decolorizer).
- Counterstain by covering the smear with Safranin for 2 minutes.
- Gently wash with DI water, blot, and observe under oil immersion.
Results
Draw and color (or indicate colors of) your Endospore stain. Label vegetative cells and endospores.
