Graham's
Aluminum Hematoxylin

Grahams alum hematoxylin is a modification of Gill's alum hematoxylin, and was designed for routine staining of plant tissue. It should also be satisfactory for animal tissues. It has been formulated so that small quantities may be prepared fresh, sufficient for a single Coplin jar.

Solution A Amount Function
Distilled water 750 mL Solvent
Propylene glycol 210 mL Solvent
Glacial acetic acid 20 mL Acidifier
Aluminum sulphate 17.6 g Mordant
Sodium iodate 0.2 g Oxidant
 
Solution B Amount Function
Hematoxylin 10 g Dye
Propylene glycol 100 mL Solvent

 

Compounding procedure

Solution A
Combine all the ingredients and filter when dissolved.

Solution B
Shake vigorously several times during 48 hours until the hematoxylin dissolves.

Working solution
Solution A - 49 mL
Solution B - 1 mL
Mix well.
It is ready for use when an opaque dark purple.

Method

  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for an appropriate time.
  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. Staining is progressive, application time being determined by trial (try 2 minutes).
  2. Propylene glycol replaces toxic ethylene glycol.
  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.
  4. The aluminum sulphate should be Al2(SO4)3.18H2O. Adjust the amount if different.

Reference
Graham, E. T., (1991)
A quick mix aluminum hematoxylin stain
Biotechnic and Histochemistry, pp. 279-281

 


 

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