Masson's Iron Hematoxylin
Materials
Solution A
| Material | Amount | Function |
|---|---|---|
| Ferric ammonium sulfate | 4 g | Mordant |
| Distilled water | 100 mL | Solvent |
Solution B
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 1 g | Dye |
| Distilled water | 100 mL | Solvent |
Compounding Procedure
- Make each solution separately.
- Solution B should be ripened for a minimum of one month.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into solution A for 5-10 minutes preheated to 50°C.
- Rinse with distilled water.
- Place into solution B for 5-10 minutes preheated to 50°C.
- Rinse with tap water.
- Differentiate in solution A at room temperature, controlling microscopically.
- Wash well in running tap water to blue.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei and other structures – blue to black
Notes
- This is a modification of Heidenhain’s iron hematoxylin, and differs solely in the concentration of the reagents, and the temperature at which staining is carried out.
- The stock solutions are stable for some time.
- The hematoxylin solution needs to be ripened.
- The degree of differentiation will determine which tissue components are prominent. The method can demonstrate many structures, including chromosomes, nuclear components, mitochondria and muscle striations
- The solutions may be reused, with the exception of the solution A used to differentiate, which should be fresh each time.
- Counterstaining is not recommended.
- This method is usually recommended for monochrome photography.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Gray, Peter. (1954)
The Microtomist’s Formulary and Guide.
Originally published by: The Blakiston Co.
Republished by: Robert E. Krieger Publishing Co.
Citing:
Heidenhain, M., (1892).
Festschrift Herrn A. von Kolloker zur Feier seines fünfzigjährigen medicinischen
Doktorjubiläums, p.118. Wilhelm Engellmans, Leipzig, Germany
and:
Masson, M.,, (1892).
Bulletin et mémoires de la Société anatomique de Paris, v.87. p.291.
