⑫ Staining

Berlin blue staining

Berlin blue staining stains trivalent iron and hemosiderin blue.

This method stains ferric iron blue. It is used to detect hemosiderin. This reaction is extremely sensitive and will demonstrate very small amount of iron.

Staining procedure

1	Deparaffinization	Xylene	3 changes, 10 minuntes each
2	Removal of xylene	100% ethanol	3 changes, 5 minutes each
3	Hydration	95%, 70% ethanol	5 minutes each
4	Washing	Running tap water	2 minutes
5	Rinsing	Distilled water
6	Staining	Pottasium ferrocyanide-HCl mixture	15 to 30 minutes
7	Washing	Distilled water	3 changes
8	Counterstaining	Kernechtrot	5 minutes
9	Washing	Distilled water	3 changes
10	Dehydration	70%, 95% ethanol	5 minutes each
11	Dehydration	100% ehtanol	3 changes, 5 minutes each
12	Clearing	Xylene	3 changes, 10 minuntes each
13	Coverslipping

Staining solution

Solution 1：2% potassium ferrocyanide (ferrocyan-kalium) aqueous solution
Solution 2：1% hydrochloric acid solution
Mix equal amounts of solutions 1 and 2 immediately before use.