Perls Staining Methods for Brain Iron

Perls Prussian blue method was followed for staining ferric iron: The ferric iron combines with potassium ferrocyanide to form the insoluble Prussian blue precipitate as follows:

Paraffin fixed brain sections (n = 6, in glass slides) were deparaffinized and hydrated with iron free distilled water. The sections were then transferred to a mixture of equal parts of 2% potassium ferrocyanide for Perls staining at pH6 and 2% HCL in distilled water for 20–30 min. The brain sections were then washed in distilled water. All sections were counterstained in eosin, dehydrated, and mounted with a synthetic resin medium (Meguro et al., 2005).