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Kiernan’s Eriochrome Cyanin for Nuclei

Kiernan's Eriochrome Cyanin

for Nuclei

8
steps
6
materials

This is a substitute for alum hematoxylin in the H & E stain.

Materials

Solution A

MaterialAmount
Eriochrome cyanine R1.0g
Ferric chloride, 5.6%20mL
Sulphuric acid, conc.2.5mL
Distilled waterup to 500 mL

Solution B

MaterialAmount
Hydrochloric acid, conc.5mL
Ethanol, abs.500mL
Distilled water500mL

Tissue Sample

5 µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.

Protocol

  1. Bring sections to water via xylene and ethanol.
  2. Place sections into solution A for 5 minutes.
  3. Wash with running tap water for about 30 seconds.
  4. Differentiate with solution B until nuclei are sharply stained, about 10-30 seconds.
  5. Wash with running tap water for about 5 minutes.
  6. Counterstain with eosin or another contrasting dye, as wished.
  7. Dehydrate with ethanol.
  8. 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

  1. Kiernan. J.A., (1999)
    Histological and histochemical methods: Theory and practice, Ed. 3
    Butterworth Heinemann, Oxford, UK.