Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections

• Published in: The journal of histochemistry and cytochemistry 1991-06, Vol.39 (6), p.741-748
• Main Authors: Shi, SR, Key, ME, Kalra, KL
• Format: Article
• Language: English
• Subjects: Antibodies, Monoclonal; Antigens - analysis; Biological and medical sciences; Cell structures and functions; Fixatives; Formaldehyde; Fundamental and applied biological sciences. Psychology; Humans; Immunohistochemistry - methods; Microwaves; Molecular and cellular biology; Paraffin; Sensitivity and Specificity; Staining and Labeling - methods
• ISSN: 0022-1554; 1551-5044
• DOI: 10.1177/39.6.1709656

We describe a new approach for retrieval of antigens from formalin-fixed, paraffin-embedded tissues and their subsequent staining by immunohistochemical techniques. This method of antigen retrieval is based on microwave heating of tissue sections attached to microscope slides to temperatures up to 100 degrees C in the presence of metal solutions. Among 52 monoclonal and polyclonal antibodies tested by this method, 39 antibodies demonstrated a significant increase in immunostaining, nine antibodies showed no change, and four antibodies showed reduced immunostaining. In particular, excellent immunostaining results were obtained with a monoclonal antibody to vimentin as well as several different keratin antibodies on routine formalin-fixed tissue sections after pre-treatment of the slides with this method. These results showed that after antigen retrieval: (a) enzyme predigestion of tissues could be omitted; (b) incubation times of primary antibodies could be significantly reduced, or dilutions of primary antibodies could be increased; (c) adequate staining could be achieved in long-term formalin-fixed tissues that failed to stain by conventional methods; and (d) certain antibodies which were typically unreactive with formalin-fixed tissues gave excellent staining.