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.


  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


  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


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




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