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      Three-dimensional Reconstruction of the Vascular Architecture of the Passive CLARITY-cleared Mouse Ovary

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          Abstract

          The ovary is the main organ of the female reproductive system and is essential for the production of female gametes and for controlling the endocrine system, but the complex structural relationships and three-dimensional (3D) vasculature architectures of the ovary are not well described. In order to visualize the 3D connections and architecture of blood vessels in the intact ovary, the first important step is to make the ovary optically clear. In order to avoid tissue shrinkage, we used the hydrogel fixation-based passive CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/ Immunostaining/In situ-hybridization-compatible Tissue Hydrogel) protocol method to clear an intact ovary. Immunostaining, advanced multiphoton confocal microscopy, and 3D image-reconstructions were then used for the visualization of ovarian vessels and follicular capillaries. Using this approach, we showed a significant positive correlation (P <0.01) between the length of the follicular capillaries and volume of the follicular wall.

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          Author and article information

          Journal
          J Vis Exp
          J Vis Exp
          JoVE
          Journal of Visualized Experiments : JoVE
          MyJove Corporation
          1940-087X
          2017
          10 December 2017
          : 130
          : 56141
          Affiliations
          1Department of Integrative Medicine and Neurobiology, School of Basic Medical Sciences; Institutes of Brain Science, Brain Science Collaborative Innovation Center, State Key Laboratory of Medical Neurobiology, Institute of Acupuncture and Moxibustion, Fudan Institutes of Integrative Medicine, Fudan University
          2Program of Reproductive and Stem Cell Biology, Department of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford University
          Author notes

          Correspondence to: Yi Feng at fengyi17@ 123456fudan.edu.cn

          Article
          PMC5755557 PMC5755557 5755557 56141
          10.3791/56141
          5755557
          29286393
          42a2ad09-2924-4af0-a718-b7bce363f2d1
          Copyright © 2017, Journal of Visualized Experiments
          History
          Categories
          Biomedical Engineering

          Issue 130,passive CLARITY,mouse,ovary,Biomedical Engineering,Three-dimensional reconstruction,vasculature

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