Hematoxylin Stain
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Hematoxylin Stain
Solutions
Hematoxylin
| Hematoxylin | 6.4g |
| Ammonium alum | 60g |
| Ethanol | 200ml |
| Glycerol | 160ml |
| Distilled water | 640ml |
Mix for a day then allow the solution to ripen in the dark for at least a month before using.
Acidic Alcohol
| Concentrated HCl | 4ml |
| 95% Ethanol | 396ml |
Bluing Agent
| Sodium bicarbonate | 1g |
| Distilled water | 1 l |
Staining
- Deparaffin and rehydrate slides.
- Slightly overstain the sections with hematoxylin 3-5 minutes.
- Remove excess stain in tap water for 2 minutes.
- Differentiate and restain a few seconds in acidic alcohol until sections look red.
- Rinse in tap water.
- Blue in bicarbonate until nuclei stand out sharply blue for 2 minutes.
- Rinse in running tap water.
- Dehydrate and clear, coverslip with Cytoseal.
