8/6/2019 Pressure Cooking Antigen Retrieval

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Chapter 2 Pressure Cooker Heating

IntroductionPressure cookers set to approximately 103 kPa/15 psi will achieve a temperature ofapproximately 120 C at full pressure. Alternatively, setting at 125 C can be used forantigen retrieval. Stainless steel pressure cookers are recommended as the aluminiummodels are susceptible to corrosion by some retrieval solutions. As an alternative,individual plastic container(s) can be flled with retrieval solution and placed in reagent gradewater in the pressure cooker pan.

Materials RequiredStainless steel pressure cooker, pre erably electrically programmable

Metal or plastic slide racksRetrieval solutionSilanized Slides (Code S3003) or slides coated with other suitable adhesivesTris-Bu ered Saline (Code S3001)Incubation container (optional)Personal protective equipment

ProtocolIt is recommended to wear a sa ety ace shield and insulated gloves.

1. Depara fnize and rehydrate tissue sections.2. Fill the pressure cooker with enough retrieval solution to cover slides. Alternatively,

fll individual plastic container(s) with retrieval solution and add at least 500 mL oreagent grade water to pressure cooker chamber.

3. Bring contents to near boiling point, place racked slides into retrieval solution, sealthe pressure cooker, and again bring the solution to a boil. For programmablepressure cookers, set target temperature and heating time, place racked slides inretrieval solution, seal the pressure cooker, and begin antigen retrieval procedurerom room temperature.

4. Boil or 30 seconds to 5 minutes, allow the pressure cooker to cool or 20 minutesprior to opening. (Note: Vent any residual pressure be ore opening). Openprogrammable pressure cooker when antigen retrieval procedure is completed.

5. Trans er slides to room temperature Tris-Bu ered Saline. When removing the slidesrom the container it is very important that the slides do not dry out.

6. Proceed with IHC staining procedure.

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