Krutsay's
Alum & Iron Hematoxylin

Krutsay's alum hematoxylin gives very highly selective staining of nuclei. It may also be easily converted into an iron hematoxylin for use as an acid resistant nuclear stain.

Alum hematoxylin Amount Function
Hematoxylin 1 g Dye
Potassium alum 50 g Mordant
Distilled water 1 L Solvent
Hydrochloric acid 5 mL Acidifier
Sodium iodate 0.2 g Oxidant
 
Iron conversion Amount Function
Iron alum 2 g Mordant
Hydrochloric acid 0.5 mL Acidifer
Distilled water 100 mL Solvent

 

Compounding procedure
Mix the reagents under Alum hematoxylin together.
Bring to the boil
Cool and filter.
The solution may be used immediately.

Conversion to iron hematoxylin
Mix the reagents under Iron conversion together, and dissolve.
For use mix the following:–
Alum hematoxylin solution – 25 mL
Iron conversion solution – 2 mL

Method

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

Expected results

Notes

  1. This is a progressive solution and is highly selective for nuclei.
  2. The iron alum is (FeNH4(SO4)2.12 H2O)
  3. 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
Humason, G. L.
Animal Tissue Techniques.
W. H. Freeman and Co., San Francisco, CA, USA

 


 

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