To detect ferric (FE3+) iron, potassium ferrocyanide is used instead in the Perls' Prussian blue staining method.
The original stain formula known historically (1867) as "Perls' Prussian blue" after its inventor German pathologist Max Perls (1843-1881), used separate solutions of potassium ferrocyanide and acid to stain tissue (these are now used combined just before staining).
Ferric iron deposits in tissue (present mostly as ferric iron within the storage protein ferritin) then react with the soluble ferrocyanide in the stain, to form insoluble Prussian blue dye (a complex hydrated ferric ferrocyanide substance) in situ. They are then visualisable microscopically as blue or purple deposits, within cells.
Prussian blue was the first modern synthetic pigment. It is prepared as a very fine colloidal dispersion because the compound is not soluble in water. It contains variable amounts of other ions and its appearance depends sensitively on the size of the colloidal particles. The pigments is used in paints and it is the traditional "blue" in Japanese woodblock prints.
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