Make each solution seperately.
Solution B should be ripened for a minimum of one month.
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.
Nuclei and other structures – blue to black
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.
Reference 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
Wilhelm Engellmans, Leipzig, Germany
and:– Masson, M.,, (1892). Bulletin et mémoires de la Société anatomique de Paris,v.87. p.291.