The action of sex steroids on the growth and differentiation of target tissues requires the presence of specific intracellular receptors. We compared the distribution of cells containing estrogen receptor (ER) and/or progesterone receptor (PR) in rabbit uterus by immunohistochemistry using monoclonal antibodies directed against these receptors. Initial experiments using serial cryostat sections surprisingly revealed the intensity of staining for ER to be inversely proportional to that of PR, as follows: ER, luminal and glandular epithelium greater than myometrium greater than stroma; PR, stroma greater than myometrium greater than glands greater than luminal epithelium. Localization was strictly confined to the nuclei of target cells. Single and dual immunofluorescent labeling of ER and PR in cryostat sections was accomplished using fluorochromes with differing emission spectra. Individual fields of dual labeled sections were examined for red [phycoerythrin (ER)] and green [fluorescein (PR)] fluorescence, with the same distribution as noted by single antibody immunohistochemistry. Myometrial nuclei displayed fluorescence of equivalent relative intensity for both antibodies. Further, sequential exposure photomicrography (exposure first in the spectrum of phycoerythrin emission, followed by exposure in the spectrum of fluorescein emission) revealed the presence of occasional stromal cells staining only for PR and some luminal cells staining only for ER. This differential distribution of ER and PR within various cell populations of rabbit is a novel observation and challenges current concepts of receptor regulation. Dual immunofluorescent localization of both ER and PR within individual cells provides a unique perspective from which to investigate the interactive influences of these sex steroid receptors at the cellular level.