Leung & Gibbon's Alcian Yellow-Toluidine Blue
for Helicobacter
Materials
Solution A
| Material | Amount | |
|---|---|---|
| Periodic acid | 1 | g |
| Distilled water | 100 | mL |
Solution B
| Material | Amount | |
|---|---|---|
| Sodium metabisulfite | 5 | g |
| Hydrochloric acid, 1N | 1 | mL |
| Distilled water | 100 | mL |
Solution C
| Material | Amount | |
|---|---|---|
| Alcian yellow | 1 | g |
| Ethanol, 50% | 100 | mL |
| Acetic acid, glacial | 3 | mL |
Filter before use.
Solution D
| Material | Amount | |
|---|---|---|
| Toluidine blue 1% aqueous | 0.5 | mL |
| Sodium hydroxide, 3% aqu. | 2 | drops |
| Distilled water | 50 | mL |
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives have not been evaluated.
Protocol
- Dewax and hydrate sections.
- Oxidize in solution A for 10 minutes.
- Wash well with water.
- Treat with solution B for 5 minutes.
- Wash in water for 2 minutes.
- Stain in solution C for 5 minutes.
- Wash well with water.
- Stain in freshly prepared solution D for 3 minutes.
- Wash well with water.
- Blot dry.
- Dehydrate, clear and mount.
Expected Results
- Helicobacter – blue
- Mucin – yellow
- Background – pale blue
Notes
- This is most useful as a screening method.
- Alcian yellow is a monoazo dye that stains similarly to alcian blue. After oxidation, gastric mucin is stained yellow and cannot later be replaced by toluidine blue. The final result is blue bacteria on a yellow background.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Leung, K., & Gibbon, K.J., (1996)
A rapid staining method for Helicobacter pylori in gastric biopsies
Journal of Histotechnology, v. 19, pp. 131
