Tirmann Schmelzer Turnbull's Blue
for Ferrous Iron
Iron deposits in the body are usually ferric compounds such as hemosiderin. Occasionally ferrous iron deposits are encountered, sometimes from mining activity or foreign bodies. These iron deposits may be demonstrated with Tirmann Schmelzers’s Turnbull’s blue. The method is very similar to Perls’ Prussian blue, but uses potassium ferricyanide instead of ferrocyanide. The ferrous iron reacts with the potassium ferricyanide to form ferrous ferricyanide. This is an insoluble, blue compound known as Turnbull’s blue. The intensity of the colour gives some indication as to amount, but it is qualitative only.
Materials
- Neutral red
- Stock solution A
Material Amount Potassium ferricyanide 20 g Distilled water 100 mL - Stock solution B
Material Amount Hydrochloric acid, conc. 1 mL Distilled water 100 mL - Working solution
Material Amount Stock solution A 1 volume Stock solution B 1 volume
Tissue Sample
5 µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Avoid iron containing materials and jars while fixing as these may contaminate the tissue. Acid containing fixatives may remove some of the iron deposits, but apart from that most are satisfactory.
Protocol
- Bring sections to distilled water with xylene and ethanol.
- Place into the working solution for 15 minutes.
- Rinse with distilled water.
- Wash well with tap water.
- Stain with neutral red for one minute.
- Rinse well with tap water.
- Dehydrate with ethanol.
- Clear with xylene.
Expected Results
- Ferrous iron – blue
- Nuclei – red
Notes
- The working solution should be made immediately before use.
- Avoid washing with tap water before placing into the working solution, as rust in the water or tap fixtures could cause false positive staining.
- Wash well at steps 3 and 4, as traces of iron will form a granular red deposit with neutral red, or use nuclear fast red.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Culling C.F.A., (1974)
Handbook of histopathological and histochemical techniques Ed. 3
Butterworth, London, UK.
