Gadsdon's
Alum Hematoxylin

This formula is by Prof. Derek Gadsdon at the Sheffield Children's Hospital, UK.

Ingredient	Amount	Function
Hematoxylin	5.5 g	Dye
Potassium alum	60 g	Mordant
Distilled water	610 mL	Solvent
100% ethanol	100 mL	Solvent
Glycerol	300 mL	Stabiliser
Sodium iodate	0.5 g	Oxidant
Glacial acetic acid	20 mL	Acidifier

 

Compounding procedure
Dissolve the hematoxylin in 100 mL ethanol.
Dissolve the Alum in 600 mL water.
Dissolve the sodium iodate in 10 mL water.
Combine the hematoxylin and Alum solutions. Mix well.
Add the sodium iodate solution. Mix well.
Add the acetic acid. Mix well.
Leave overnight.
Add the glycerol. Mix well.
Filter before use.
The solution may be used immediately.
It is stable at room temperature for at least 6 months.

Method

1. Bring sections to water with xylene and ethanol.
2. Place into the staining solution for the appropriate time.
3. Rinse well with water.
4. Differentiate with acid ethanol if using regressively.
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

• Nuclei  –  blue
• Background  –  as counterstain or unstained

Notes

1. This solution may be used regressively or progressively.
   Regressively, staining time is 6 minutes.
   Progressively, staining time is 3 minutes.
   Frozen sections will stain in 1 minute.
2. As with many strong formulations mucins may stain blue.
3. Acid ethanol is 0.5% - 1% hydrochloric acid in 70% ethanol.
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
Histopathology laboratory, Red Cross Children's Hospital.
Gadsdon's Haematoxylin Method
Histonet communication, Nov, 1996

Jim Elsam, HTEQA Services.
Gadsdon's Hx
Histonet communication, Nov, 1996

 

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