Kiernan's Eriochrome Cyanin
for Nuclei

Substitute for alum hematoxylin in the H&E stain

Solutions

Solution A
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
Hydrochloric acid, conc.	5	mL
Ethanol, abs.	500	mL
Distilled water	500	mL

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

Method

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 ethanols.
8. Clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  blue
• Background  –  as counterstained

Notes

1. Solution A is stable for years.
2. Solution A is also used for Kiernan's myelin staining variant.
3. Kiernan also describes a two colour staining variant.

 

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

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