Polo-like kinase 1 (Plk1) has been reported to localize to the spindle midzone during meiosis in mouse oocytes. However, it has not been reported in human oocytes. In this study, the interaction of the meiotic structures and chromosome segregation in mouse and human oocytes were studied by time-lapse differential interference contrast microscopy. Using immunocytochemical studies, the localization of polo-like kinase 1 and its association with microtubules were examined during the extrusion of first and second polar bodies. It was found that Plk1 was localized in the spindle midzone in human oocytes at anaphase I and telophase I. Also, three-dimensional confocal laser microscopy showed that the meiotic spindle midzone contained numerous dot-like particles that were stained by anti-Plk1 antibody. These particles were aligned in the plane of the meiotic midzone in mouse and human oocytes.