General Procedure for Antigen Retrieval Using Microwave Heating

GENERAL PROCEDURE FOR ANTIGEN RETRIEVAL USING MICROWAVE HEATING 

Fix the tissue without delay for4-6hr in 4% buffered formalin the longer the tissue remains in the fixative, the lesser the chances of epitope retrieval (Hayat, 

Wash in several changes of PBS if available, an Autotecnicon should be used in this step and in steps 3 and 4 below. 

Cut 4-pjn-thick sections with a microtome and float them on a water bath kept at room temperature so as to stretch the sections. An ordinary glass slide is used to transfer the sections onto another water bath kept at 58°C to further stretch the sections. Lift them by the SuperFrost slides, thus mounting them in the process. The sections are allowed to dry in an upright position in a slide holder at a temperature of 30°C. When the slide holder is full, it is transferred to a conventional oven. 

Remove the sections from the oven and deparaffinize them with three changes of 5 min each in xylene, followed by three changes of 100% ethanol, two changes of 75% ethanol, and three changes of distilled water. A wash in Tris-buffered saline (TBS) (pH 7.3) for 10 min is optional. 

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Hayat, M. A. 1999. General procedure for antigen retrieval using microwave heating. Microsc. Today 99(7) 28-30. 

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