Fig. 3

Three-dimensional image of the EGFP-positive mouse ovary. Female EGFP-transgenic mice were transcardially perfused with 4% PFA containing PI. After the isolated ovaries were subjected to CUBIC, 3D and cross-sectional images were taken by light-sheet microscopy (a, b and c) or confocal microscopy (d and e). a A 3D image of EGFP and PI signals of the whole ovary. b A sectional image within the blue square of (a). c A higher-magnification image within the white square of (b). d Preantral follicles. e Mature antral follicle. Note that EGFP fluorescence was strongly visible in the oocyte and ovarian stromal cells, but not in granulosa cells around oocytes. gr, granulosa cells; o, oocyte; th, thecal cells; epi, surface epithelial cells; str, stromal cells. Scale bars, 500 μm (a), 100 μm (b, d and e), and 50 μm (c)