Chapter 2 Pressure Cooker Heating

Introduction
Pressure cookers set to approximately 103 kPa/15 psi will achieve a temperature of approximately 120 C at full pressure. Alternatively, setting at 125 C can be used for antigen retrieval. Stainless steel pressure cookers are recommended as the aluminium models are susceptible to corrosion by some retrieval solutions. As an alternative, individual plastic container(s) can be filled with retrieval solution and placed in reagent grade water in the pressure cooker pan.

Materials Required
Stainless steel pressure cooker, preferably electrically programmable Metal or plastic slide racks Retrieval solution Silanized Slides (Code S3003) or slides coated with other suitable adhesives Tris-Buffered Saline (Code S3001) Incubation container (optional) Personal protective equipment

Protocol
It is recommended to wear a safety face shield and insulated gloves. 1. Deparaffinize and rehydrate tissue sections. 2. Fill the pressure cooker with enough retrieval solution to cover slides. Alternatively, fill individual plastic container(s) with retrieval solution and add at least 500 mL of reagent grade water to pressure cooker chamber. 3. Bring contents to near boiling point, place racked slides into retrieval solution, seal the pressure cooker, and again bring the solution to a boil. For programmable pressure cookers, set target temperature and heating time, place racked slides in retrieval solution, seal the pressure cooker, and begin antigen retrieval procedure from room temperature. 4. Boil for 30 seconds to 5 minutes, allow the pressure cooker to cool for 20 minutes prior to opening. (Note: Vent any residual pressure before opening). Open programmable pressure cooker when antigen retrieval procedure is completed. 5. Transfer slides to room temperature Tris-Buffered Saline. When removing the slides from the container it is very important that the slides do not dry out. 6. Proceed with IHC staining procedure.