Goldman's Iron Hematoxylin
Materials
Solution A
| Material | Amount | Function |
|---|---|---|
| Ferric ammonium sulfate | 4 g | Mordant |
| Distilled water | 100 mL | Solvent |
| Acetic acid, glacial | 1 mL | Acidifier |
| Sulfuric acid | 0.12 mL | Acidifier |
Solution B
| Material | Amount | Function |
|---|---|---|
| Picric acid, saturated aqueous | 100 mL | Dye and acid |
| Sulfuric acid | 0.1 mL | Acidifier |
Solution C
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 0.5 g | Dye |
| Distilled water | 100 mL | Solvent |
Solution D
| Material | Amount | Function |
|---|---|---|
| 70% ethanol | 100 mL | Solvent |
| Lithium carbonate, saturated aqueous | 5 drops | Base |
Compounding procedures
- Make each solution separately.
- The aqueous hematoxylin should be ripened before use.
Protocol
- Bring sections to water, removing mercury pigment if necessary.
- Place into solution A for 30 minutes to 24 hours.
- Wash in running tap water for 10 minutes.
- Place in solution B for 3 hours or longer.
- Wash in running tap water for 15 minutes.
- Place in solution C for 1 hour.
- Wash in running tap water for 15 minutes.
- Blue with solution D.
- Counterstain if desired.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – black
- Basophil cytoplasm – grey
- Background – as counterstain or unstained
Notes
- The stock solutions are stable for some time.
- The technique was originally intended for the demonstration of protozoa.
- The method was designed for paraffin sections of material fixed with formalin variants, Bouin’s or Zenker’s fluids.
- Overstaining occurs only if sections are left in hematoxylin for several hours.
- Bouin fixed tissue is not as intensely stained as with other fixatives.
- The time in picric acid (solution B) is necessary, reducing it causes overstaining.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Lillie, R.D., (1954)
Histopathologic technique and practical histochemistry Ed.2
Blakiston, New York, USA.
Citing:
Goldman, (1951)
American Journal of Clinical Pathology,
v.21, p.198
