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. 2020 Jan 24;102(5):1080–1089. doi: 10.1093/biolre/ioaa012

Figure 2.

Figure 2

Combined iDISCO and CUBIC clearing for in toto imaging of the adult mouse ovary. (A) Schematic diagram of the experimental workflow for optical clearing and fluorescent immunostaining of intact mouse ovaries using the combination of iDISCO and CUBIC methods. (B) Photograph of ovaries cleared following the iDISCO protocol in DBE, visualized with halogen (top) or UV (bottom) light. White arrows point to cleared ovaries. (C) Photomicrograph of uncleared (top) versus iDISCO + CUBIC-cleared (bottom) 2-month-old ovaries (scale bar, 1 mm). (D) Optical sections from the surface (left) and center (right) z-planes of LSFM images of iDISCO + CUBIC-cleared ovary from a 2-month-old mouse labeled with antibodies against interstitial marker αSMA (green) and growing follicle marker AMH (red). Insets are magnified views from the areas outlined in top row images (scale bars, 300 μm). (E) 3D render generated from LSFM images of iDISCO + CUBIC-cleared ovary shown in D. The inset is a magnified view of a single growing follicle from the area outlined in the top row image (scale bar, 50 μm).