Microwave stabilization enhances immunocytochemical detection of estrogen receptor in frozen sections of macaque oviduct

Ov Slayden, T. Koji, R. M. Brenner

    Research output: Contribution to journalArticle

    42 Citations (Scopus)

    Abstract

    We have found that microwave (MW) stabilization greatly improves detection of the estrogen receptor (ER) in frozen sections of rhesus monkey oviduct by immunocytochemistry (ICC). Fresh samples of fimbriae were MW-irradiated, frozen, and then cryosectioned. The frozen sections were also MW-treated and then fixed in a paraformaldehyde-based fixative before ICC processing. A parallel set of samples from each monkey were frozen, sectioned and processed for ICC without any MW treatment. MW stabilization clearly increased immunostaining intensity with either of two ER-specific monoclonal antibodies, namely, H222 and 1D5. The greatest increase was noted in tissues collected from spayed or progesterone-treated animals. An antibody dilution series indicated that MW stabilization increased the sensitivity approximately 20- to 40-fold. In addition, we incubated spayed macaque fimbriae at 4 C in the presence of 10 nM [3H]Moxestrol and then either froze the tissues immediately (non-MW) or treated them with MW. Slide-mounted cryosections of non-MW and MW-treated tissue were then incubated with either a Tris-EDTA buffer (low salt) or the same buffer containing 4 M KCl (high salt). The quantity of [3H]Moxestrol-occupied ER extracted from the frozen sections by each buffer was determined by a sucrose gradient shift assay. The low salt buffer extracted significantly more radiolabeled ER from non-MW sections than from MW-treated sections (P <0.01), whereas the high salt buffer extracted equal amounts of ER from both the MW-treated and non-MW sections. MW-irradiation enhanced ICC detectability of ER in frozen sections by greatly reducing the amount of ER extracted during the various washes used during normal ICC processing.

    Original languageEnglish (US)
    Pages (from-to)4012-4021
    Number of pages10
    JournalEndocrinology
    Volume136
    Issue number9
    StatePublished - 1995

    Fingerprint

    Oviducts
    Frozen Sections
    Macaca
    Microwaves
    Estrogen Receptors
    moxestrol
    Immunohistochemistry
    Buffers
    Salts
    Fixatives
    Tromethamine
    Macaca mulatta
    Edetic Acid
    Haplorhini
    Progesterone
    Sucrose
    Monoclonal Antibodies

    ASJC Scopus subject areas

    • Endocrinology
    • Endocrinology, Diabetes and Metabolism

    Cite this

    Microwave stabilization enhances immunocytochemical detection of estrogen receptor in frozen sections of macaque oviduct. / Slayden, Ov; Koji, T.; Brenner, R. M.

    In: Endocrinology, Vol. 136, No. 9, 1995, p. 4012-4021.

    Research output: Contribution to journalArticle

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