Kiernan's Eriochrome Cyanin
with two colours


Solution A
Eriochrome cyanine R 1.0 g
Ferric chloride, 5.6% 2.5 mL
Sulphuric acid, conc. 2.5 mL
Distilled water up to 500 mL

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


  1. Bring sections to water via xylene and ethanol.
  2. Place sections into solution A for 3 minutes.
  3. Rinse with distilled water three times for about 20 seconds each time.
  4. Dehydrate with ethanols.
  5. Clear with xylene and mount with a resinous medium.

Expected results


  1. Solution A is stable for years.
  2. Solution A must have a pH of 1.5 for the two colour effect. Check periodically, and adjust with 1M sulphuric or hydrochloric acids, or sodium hydroxide.
  3. Kiernan's also describes regressive nuclear staining and myelin staining variants.
  4. Eriochrome cyanine R is also known as mordant blue 3 and solochrome cyanine R


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



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