Horneyold's
Alum Hematoxylin

Hematoxylin solution Amount Function
Hematoxylin 0.7 g Dye
100% ethanol 20 mL Solvent
 
Alum solution Amount Function
Ammonium alum 0.35 g Mordant
Distilled water 60 mL Solvent
 
Tincture of iodine Amount Function
Iodine 70 g Oxidant
Potassium iodide 50 g
Distilled water 50 mL Solvent
100% ethanol 950 mL Solvent

 

Compounding procedure

Tincture of iodine
Mix the dry iodine and potassium iodide together.
Add the water and mix well to dissolve.
Add the ethanol and mix well.

Staining solution
Dissolve the hematoxylin in the ethanol and the alum in the water.
Combine the two solutions and mix well.
Leave in the light by a window for 3-4 days.
Add 20 drops tincture of iodine.

Method

  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for 5-10 minutes.
  3. Rinse well with water.
  4. Differentiate with acetic ethanol.
  5. Rinse with water and blue.
  6. Rinse well with water.
  7. Counterstain if desired.
  8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

Notes

  1. Horneyold's formula is a modification of Böhmer's alum hematoxylin.
  2. This solution is regressive, and is recommended for osmium fixed tissue.
  3. Acetic ethanol is 70% ethanol containing glacial acetic acid. The amount of acetic acid to add was not given, but 0.5%-1% should suffice.
  4. Blueing is done with alkaline solutions such as hard tap water, Scott’s tap water substitute, 0.1% ammonia water, 1% aqueous sodium acetate, 0.5% aqueous lithium carbonate etc.

 

Reference
Bolles Lee, A.. Edited by Gatenby, J.B. and Beams, H.W., (1950)
The Microtomist's Vade-Mecum. 11 ed.,
Churchill, London, UK.

Lange, N.A., (1966)
Lange's Handbook of Chemistry, Revised 10th ed.,
McGraw-Hill.

 


 

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