Three-dimensional Reconstruction of the Vascular Architecture of the Passive CLARITY-cleared Mouse Ovary
Three-dimensional Reconstruction of the Vascular Architecture of the Passive CLARITY-cleared Mouse Ovary
复制标题
被动 CLARITY 清除小鼠卵巢血管结构的三维重建
DOI：
10.3791/56141
复制
发表时间：
2017-12-01
影响因子：
1.2
通讯作者：
Feng, Yi
中科院分区：
文献类型：
--
作者：
Hu, Wei;Tamadon, Amin;Feng, Yi
卵巢是女性生殖系统的主要器官，对于产生雌性配子和控制内分泌系统是必不可少的，但卵巢的复杂结构关系和三维（3D）脉管系统结构还没有得到很好的描述。为了可视化完整卵巢中血管的3D连接和结构，第一个重要步骤是使卵巢光学清晰。为了避免组织收缩，我们使用了基于水凝胶固定的被动免疫学（透明脂质交换丙烯酰胺杂交刚性成像/免疫染色/原位杂交相容组织水凝胶）方案方法来清除完整的卵巢。免疫染色，先进的多光子共聚焦显微镜，三维图像重建，然后用于卵巢血管和卵泡毛细血管的可视化。结果表明，卵泡毛细血管长度与卵泡壁体积呈显著正相关（P <0.01）。
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.