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Unmasking using a Steamer

A HTAU/HIER method using a steamer

A slow but still effective way to heat sections to the required temperature for unmasking. This method may take twice as long as microwave methods for comparable staining.

  1. Cut & mount sections on coated slides - VECTABOND , Poly L-Lysine or APES coatings are recommended.
  2. Deparaffinize sections and rehydrate to distilled water
  3. Fill steamer reservoir with distilled water
  4. Add a vessel with the retrieval solution to the steaming chamber.
  5. Heat until the system & retrieval buffer is equilibrated to 95°C (about 75 minutes for 200ml retrieval solution)
  6. Add sections & steam for 20-40 minutes
  7. Remove sections and retrieval buffer from the steamer & allow to cool for 20 minutes at room temperature. Be careful to avoid scalds from the steam.
  8. Rinse in distilled water.
  9. Continue with immunohistochemistry protocol.

Monitor the steamer reservoir during use to ensure it does not boil dry.


Commercially available high & low pH antigen unmasking solutions are suitable for this method.

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