New approaches to the estimation of endometrial dysfunction

Abstract

Introduction. The application of modern methods for assessing the morphofunctional state of the endometrium to verify and study the expression of sex steroid hormones, proinflammatory markers and markers of angiogenesis using confocal laser scanning microscopy will allow an objective study of the role of studied markers in the pathogenesis of the endometrial dysfunction. The aim of the study was to evaluate the expression of ER and PR receptors in endometrium in patients with endometrial dysfunction. Material and methods. Endometrial biopsy specimens obtained with the aid of a pile-biopsy or a scraping from the uterine cavity are used to conduct the method of immunofluorescent confocal laser scanning microscopy. It is possible to use both cryostat material and paraffin blocks to provide the immunohistochemical analysis. Monoclonal antibodies to ER (1 : 60, Dako, Denmark) and PR (1 : 50, Dako, Denmark) are used as primary antibodies, antibodies conjugated with fluorochrome Alexa Fluor 647 (1 : 1000, Abcam, England) are used as secondary antibodies). Hoechst 33258 (Sigma, USA) is used for staining of cell nuclei. Results. A method of confocal laser scanning microscopy makes it possible to conduct qualitative and quantitative evaluation of the studied markers in different structures of the endometrium.