Prussian Blue Iron Stain

The page below is a sample from the LabCE course Technical Preparation of Bone Marrow Specimens for Histological Assessment. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

Learn more about Technical Preparation of Bone Marrow Specimens for Histological Assessment (online CE course)
Prussian Blue Iron Stain

The Prussian blue stain is commonly performed on any one or more of the following bone marrow biopsy specimen preparations:
  • Fat and perivascular (F-PV) smear
  • PV/myeloid-erythroid (M-E) smear
  • M-E smear
  • Formalin-fixed, paraffin-embedded (FFPE) clot section
  • FFPE core biopsy section
Laboratory staining protocols are uniquely influenced by pathologists' preferences for reading the iron stain. Iron stains are used to evaluate iron storage. An ideal smear should contain bone marrow particles for more accurate bone marrow iron storage evaluation. Bone marrow core biopsy sections are not optimal as decalcification of the sample can leach out iron.
The Prussian Blue Iron stain is used to demonstrate ferric (Fe3+) iron in tissues. The mode of action for the Prussian blue iron stain is to treat the tissue with an acidic solution (hydrochloric acid). Ferric iron that is present in the tissue will react to form Prussian blue, which is visualized as an insoluble blue pigment.
Absent iron stores in a bone marrow biopsy specimen would indicate an iron deficiency, whereas increased iron stores would indicate anemia of chronic disease due to ineffective erythropoiesis (red blood cell production).

Bone marrow biopsy demonstrating increased iron stores

Bone marrow biopsy demonstrating a lack of iron stores
Bone marrow biopsy demonstrating normal iron stores