Bensley’s
Copper Hematoxylin

Solution A	Amount	Function
Hematoxylin,	1 g	Dye
Distilled water	100 mL	Solvent
Solution B	Amount	Function
Cupric acetate,	to saturation	Mordant
Distilled water	100 mL	Solvent
Solution C	Amount	Function
Potassium chromate,	5 g	–
Distilled water	100 mL	Solvent
Solution D	Amount	Function
Weigert’s borax-ferricyanide	20 mL	Differentiator
Distilled water	80 mL	Diluent

 

Compounding procedure
Make up each solution seperately.

Method

1. Bring sections to water with xylene and ethanol.
2. Place into solution A for 5 minutes.
3. Rinse with water.
4. Dip into solution B.
5. Rinse with water.
6. Place into solution C for 2 minutes.
7. Return to solution A for 1 minute.
8. Place in solution D until differentiated.
9. Rinse well with water.
10. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

• Nuclei and other structures  –  blue-black

Note

1. After step 7 the sections should be intense black. Repeat steps 5-7 if necessary.
2. Bensley recommended prestaining with Mayers mucicarmine.
3. Best results are obtained with freshly made solutions

 

Reference
Bensley R. R. and Bensley, S. H., (1938)
Handbook of Histological and Cytological Technique.
U. Chicago Press, Chicago, USA

 

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