Kiernan's Eriochrome Cyanin
for Myelin
Materials
Solution A
| Material | Amount | |
|---|---|---|
| Eriochrome cyanine R | 1.0 | g |
| Ferric chloride, 5.6% | 20 | mL |
| Sulfuric acid, conc. | 2.5 | mL |
| Distilled water | up to 500 | mL |
Solution B
- Iron alum, 10% aqu., or
- Ferric chloride, 5.6% aqu., or
- Ferric nitrate, 7.3% aqu.
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.
Protocol
- Bring sections to water via xylene and ethanol.
- Place sections into solution A for 20 minutes.
- Wash with running tap water for about 30 seconds.
- Differentiate with one of the variants of solution B until only myelin is blue, about 5-10 minutes.
- Wash with running tap water for about 5 minutes.
- Counterstain with eosin or another contrasting dye, as wished
- Dehydrate with ethanols.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Myelin – blue
- Erythrocytes – blue
- Background – as counterstained
Notes
- Solution A is stable for years.
- Solution A is also used for Kiernan’s nuclear staining variant.
- Kiernan also describes a two colour staining variant.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Kiernan. J.A., (1999)
Histological and histochemical methods: Theory and practice, Ed. 3Butterworth Heinemann, Oxford, UK.
