Duval's Alum Hematoxylin
Materials
Original Formula
| Material | Amount | Function |
|---|---|---|
| Hematoxylin, concentrated ethanolic | 8 mL | Dye |
| Ammonium or potassium alum | a little | Mordant |
| Distilled water | 800 mL | Solvent |
Modern Formula
| Material | Amount | Function |
|---|---|---|
| Hematoxylin, saturated ethanolic | 8 mL | Dye |
| Ammonium or potassium alum | 25 g | Mordant |
| Distilled water | 800 mL | Solvent |
Compounding procedure
- Dissolve the alum in the water.
- Add the hematoxylin.
- The solution is likely progressive, although this is not stated to be so.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for a few minutes.
- Rinse with water and blue.
- Rinse well with water.
- Counterstain if desired.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – blue
- Background – as counterstain or unstained
Notes
- This solution is from the late 1800’s and is now obsolete, although the modern formula should stain satisfactorily.
- Concentrated alcoholic hematoxylin, after ripening, would have contained no more than 7% hematein. It was made by soaking logwood chips in ethanol until no more dye dissolved out, then oxidized naturally. Depending on the sample of logwood and the amount of dye it contained, more than one batch may have been necessary to saturate the ethanol.
- The type of alum was not specified, the most likely being either potassium or ammonium.
- The formula calls for adding a “little alum” to 800 mL water. Potassium alum saturates at about 14% in water, so 800 mL would contain about 112 g. I have taken just less than 25% of the maximum (i.e. 25 grams) as being a “little”.
Of course, it could be any amount between 1 and 112 grams. - The appropriate time should be determined by trial. The instructions are to use full strength for a few minutes.
- 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.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Arthur Bolles-Lee, (1885)
The Microtomist’s Vade-Mecum
Originally published by: J & A Churchill, London, England.
Republished by: Science Heritage Ltd., Lincolnwood, Illinois, USA. - Susan Budavari, Editor, (1996)
The Merck Index, Ed. 12
Merck & Co., Inc., Whitehouse Station, NJ, USA
