Table 2.
Hematoxylin and Eosin staining
| Step | Solution | Time | Remarks |
|---|---|---|---|
| Deparaffinization | Xylene (or Histo-clear) | 6 min | |
| Xylene (or Histo-clear) | 6 min | ||
| 100% Ethanol | 3 min | ||
| 100% Ethanol | 3 min | ||
| 95% Ethanol | 3 min | ||
| 80% Ethanol | 3 min | ||
| Distilled water | 6 min | ||
| Hematoxylin staining | Hematoxylin solution | 3–5 min | |
| Washing | Tap water | 1 min | |
| Tap water | 1 min | ||
| Tap water | 2 min | ||
| Coloring | 0.1% Sodium bicarbonate solution | 1 min | Do not agitate |
| Washing | Tap water | 1 min | |
| Distilled water | 2 min | ||
| Eosin staining | Eosin Y solution | 30–45 sec | |
| Washing | 95% Ethanol | 2 min | |
| 95% Ethanol | 2 min | ||
| 95% Ethanol | 2 min | ||
| Dehydration | 100% Ethanol | 3 min | Use solvents dehydrated by molecular sieve |
| 100% Ethanol | 3 min | ||
| Xylene (or Histo-clear) | 10 min | ||
| Xylene (or Histo-clear) | >10 min |