A comparison of microwave heating and proteolytic pretreatment antigen retrieval techniques in formalin fixed, paraffin embedded tissues.
Antigen retrieval (AR) is a technique that re-exposes epitopes in formalin fixed, paraffin embedded sections and makes them detectable by immunohistochemistry. We compared the effects of two AR procedures, enzyme digestion and microwave heating, on immunostaining of vimentin and desmin in formalin fixed, paraffin embedded tissues. Our results showed that AR is necessary for vimentin and desmin immunostaining in tissues fixed in formalin for more than 48 h. With prolonged fixation times, microwave heating showed better results than enzyme digestion for AR. The same results were obtained using 1% zinc sulfate or Citra Plus solution as retrieval solutions for microwave heating. We recommend microwave heating for AR, because it is easier to use and produces better results compared to enzyme treatment.
Kahveci
Z
Z
Department of Histology and Embryology, Uludag University, Faculty of Medicine, Bursa, Turkey. zeynep2@uludag.edu.tr
Minbay
F Z
FZ
Noyan
S
S
Cavusoglu
I
I
D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000378:metabolism; D001011:Aorta / Q000166:cytology / Q000378:metabolism; D046508:Culture Techniques; D003893:Desmin / Q000378:metabolism*; D005557:Formaldehyde; D006361:Heating; D007150:Immunohistochemistry / Q000379:methods*; D007421:Intestine, Small / Q000166:cytology / Q000378:metabolism; D008872:Microwaves; D016612:Paraffin Embedding / Q000379:methods*; D010447:Peptide Hydrolases; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling / Q000379:methods; D016707:Tissue Fixation / Q000379:methods*; D014746:Vimentin / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000941:Antigens; D003893:Desmin; D014746:Vimentin; D005557:Formaldehyde; D010447:Peptide Hydrolases