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

1DeparaffinizationXylene3 changes, 10 minuntes each
2Removal of xylene100% ethanol3 changes, 5 minutes each
3Hydration95%, 70% ethanol5 minutes each
4WashingRunning tap water2 minutes
5RinsingDistilled water 
6StainingPottasium ferrocyanide-HCl mixture15 to 30 minutes
7WashingDistilled water3 changes
8CounterstainingKernechtrot5 minutes
9WashingDistilled water3 changes
10Dehydration70%, 95% ethanol5 minutes each
11Dehydration100% ehtanol3 changes, 5 minutes each
12ClearingXylene3 changes, 10 minuntes each

Staining solution