Microwave Antigen Retrieval Procedure

Deparaffinize and hydrate to slides to distilled water using standard immunohistochemical protocol.

Obtain a clean Tissue Tek™ plastic staining dish.

Fill a Tissue Tek™ plastic slide holder (24 slide capacity) with the hydrated slides. If there are less than 24 slides, double space the sample slides and fill the remaining slots with blank slides. 

Place the full slide rack in the plastic staining dish. Fill with 10mM sodium citrate buffer pH 6.0 up to the staining line on the dish.

Place the staining dish cover loosely (rotate the lid so that there are two small slits, one at end to act as vents so that steam can escape) over the top of the dish to prevent excess evaporation during the microwave process. If the lid tightly secured to the staining dish during the microwave process steam may build-up causing an explosion.

Place the plastic staining dish with the slides immersed in citrate buffer in the 1100w microwave located in the histology lab. Make sure there are paper towels layering the bottom of the microwave oven to catch any boil-over.

Note: For ER/PR stains use Citra Plus buffer (Biogenex).

Microwave the slides on 100% power for 2 minutes.  

Check the slides for excessive evaporation and add distilled water if necessary.

Microwave the slides using 20% power for 18 minutes.

Remove the staining dish from the microwave and remove the lid. Allow the slides to cool for 30 minutes at room temp.

Rinse the slides in distilled water and proceed with the immunohistochemical staining procedure as usual.