Kiernan's Eriochrome Cyanin
for Myelin

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
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.

Method

1. Bring sections to water via xylene and ethanol.
2. Place sections into solution A for 20 minutes.
3. Wash with running tap water for about 30 seconds.
4. Differentiate with one of the solutions C variants until only myelin is blue, about 5-10 minutes.
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

• Myelin  –  blue
• Erythrocytes  –  blue
• Background  –  as counterstained

Notes

1. Solution A is stable for years.
2. Solution A is also used for Kiernan's nuclear 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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