This formulation differs from most others in that it is made from three stock solutions. It contains no added acid, relying on the high alum:dye ratio for it's progressive staining. Consequently, the carry over of tap water fairly rapidly causes both dilution and raises the pH. Due to these, it has a shorter working life than many other formulations. It was designed to be quickly prepared in laboratories that stain large numbers of slides and prefer to use fresh solution daily, but which would like to avoid the poor background staining obtained with formulations that incorporate an acid to accentuate nuclei.
|Potassium alum||2 kg||Mordant|
|Distilled water||19.8 L||Solvent|
|Strong formalin||200 mL||Preservative|
|Strong formalin||200 mL||Solvent|
|Sodium iodate||4 g||Oxidant|
|Distilled water||200 mL||Solvent|
Dissolve the alum into 4 litres distilled water, using heat.
Add the balance of the water and the formalin.
Stock hematoxylin and stock oxidiser
Dissolve each constituent into the appropriate solvent.
Place 400 mL of the stock mordant into a 2 Litre flask.
Add 20 mL stock hematoxylin and 20 mL stock oxidiser.
Under a fume hood, bring to the boil, then remove immediately.
Add 1600 mL stock mordant.
The solution may be used immediately.
The formulated solution is stable for about 2 months at room temperature.
The stock solutions are stable at least 6 months at room temperature.
The solution may also be made in smaller volumes according to a more traditional regimen:–
|Potassium alum||100 g||Mordant|
|Distilled water||1 L||Solvent|
|Sodium iodate||0.2 g||Oxidant|
Combine the ingredients.
Bring to the boil and immediately remove from heat.
Cool to room temperature.
The solution may be used immediately it has cooled.
Slidders, W. (1988)
Preparation of alum hematoxylin.
Medical Laboratory Sciences, v 45, pp. 405.