An easy method for the removal of Epon resin from semi-thin sections. Application of the avidin-biotin technique.
A simple procedure is described for removing Epon resin from semi-thin 1 micron sections, which permits excellent postembedding immunohistochemical staining (avidin-biotin complex technique). The procedure was developed for the detection of growth hormone and prolactin in bovine adenohypophysis fixed with 2% paraformaldehyde and 0.5% glutaraldehyde in 0.1 M sodium cacodylate buffer pH 7.4-7.6. The results indicate that the removal of the epoxy embedding medium prior to the application of the immunohistochemical reagents was essential for the successful localization of the antigenic determinants of the two hormones. The immunocytochemical reactivity was obtained only after treating the sections with a solution of potassium hydroxide in a mixture of absolute methyl alcohol and propylene oxide (Maxwell's solution). An enhanced immunoreactivity was obtained when this treatment was followed by an additional treatment with either 4% hydrogen peroxide or a saturated aqueous solution of sodium metaperiodate. Because of the easy preparation of the Epon removal solution and the good structural preservation without damage to the antigenic determinants, Maxwell's solution is suggested as a good etching agent which can be used in immunohistochemical studies on semi-thin sections with excellent results.
Vidal
S
S
Department of Anatomy and Histology, University of Santiago, Faculty of Veterinary Sciences, Lugo, Spain.
Lombardero
M
M
Sánchez
P
P
Román
A
A
Moya
L
L
D000818:Animals; D001360:Avidin; D001710:Biotin; D002417:Cattle; D004853:Epoxy Resins; D013006:Growth Hormone / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods*; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000378:metabolism; D016613:Plastic Embedding / Q000379:methods*; D011388:Prolactin / Q000378:metabolism; D013194:Staining and Labeling
D004853:Epoxy Resins; D001360:Avidin; D001710:Biotin; D011388:Prolactin; D013006:Growth Hormone