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Iron Stain

Reagent Preparation

Working Iron Stain solution is prepared by mixing equal volumes of Potassium Ferrocyanide solution and Hydrochloric Acid solution. Use once and then discard. Working Pararosaniline solution is prepared by adding 1 mL of Pararosaniline solution to 50 mL water. Prepare fresh daily. Use once and discard.

Procedure for tissue sections

  1. Deparaffinize tissue sections and hydrate to deionized water.
  2. Place slides in working Iron Stain solution for 10 minutes.
  3. Rinse in deionized water.
  4. Stain in working Pararosaniline solution for 3-5 minutes.
  5. Rinse in Deionized water.
  6. Rapidly dehydrate in alcohol, clear in xylene and mount.

Procedure for blood or bone marrow films

  1. Place fixed films in working Iron Staining solution for 10 minutes.
  2. Rinse in deionized water.
  3. Counterstain 5 minutes in working Pararosaniline solution.
  4. Rinse in deionized water and air dry.


Tissue Sections

Iron Pigment – Bright blue
Nuclei – Red
Cytoplasm – Light pink

Blood or bone marrow films

Sideroblasts: These are nucleated erythrocytes containing at least one small blue granule. If the blue granules surround the nucleus, the cell is a ringed sideroblast.
Siderocytes: These are non-nucleated erythrocytes containing at least one blue granule.
Reticuloendothelial Iron: Usually seen as blue particles on the marrow film or as blue particles in the cytoplasm or phagocytic cells.

Iron Stain


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