Perl’s prussian blue staining technique for hemosiderin

Hemosiderin is present in tissues as intracellular pigment. It contains iron in the form of ferric hydroxide that is bond to a protein framework.

Principle

The reaction occurs with the treatment of tissue sections with acid ferrocyanide solution. Any ferric ion (Fe3+) in the tissue combines with ferrocyanide and results in the formaion of a bright blue pigment called “prussian blue” or ferric ferrocyanide.

Fixation

Avoid the use of acid fixatives. Chromates will also interfare with the preservation of iron.

Staining solution

  • 1% aqueous potassium ferrocyanide = 20 ml
  • 2% aqueous hydrochloric acid = 20 ml
  • Mix both

Procedure

  1. Deparaffinize and bring the sections to water.
  2. Treat the sections with freshly prepared acid ferrocyanide solution for 10-30 minutes.
  3. Wash well in distilled water.
  4. Lightly stain the nuclei with 0.5% aqueous neutral red or 0.1% nuclear fast red.
  5. Wash rapidly in distilled water.
  6. Dehydrate, clear and mount.

Result and Interpretation

pearl-prussian-blue-staining-hemosiderin

  • Ferric ion = blue
  • Nuclei = red
  • Background = pink
Perl’s prussian blue staining technique for hemosiderin
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