Kiernan's Eriochrome Cyanin
for Nuclei
This is a substitute for alum hematoxylin in the H & E stain.
Materials
Solution A
Material | Amount | |
---|---|---|
Eriochrome cyanine R | 1.0 | g |
Ferric chloride, 5.6% | 20 | mL |
Sulphuric acid, conc. | 2.5 | mL |
Distilled water | up to 500 mL |
Solution B
Material | Amount | |
---|---|---|
Hydrochloric acid, conc. | 5 | mL |
Ethanol, abs. | 500 | mL |
Distilled water | 500 | mL |
Tissue Sample
5 µ 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 5 minutes.
- Wash with running tap water for about 30 seconds.
- Differentiate with solution B until nuclei are sharply stained, about 10-30 seconds.
- Wash with running tap water for about 5 minutes.
- Counterstain with eosin or another contrasting dye, as wished.
- Dehydrate with ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – blue
- Background – as counterstained
Notes
- Solution A is stable for years.
- Solution A is also used for Kiernan’s myelin staining variant.
- Kiernan also describes a two colour staining variant.
- Eriochrome cyanine R is also known as mordant blue 3 and solochrome cyanine R.
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. 3
Butterworth Heinemann, Oxford, UK.