Willows Jake W, Blaszkiewicz Magdalena, Lamore Amy, Borer Samuel, Dubois Amanda L, Garner Emma, Breeding William P, Tilbury Karissa B, Khalil Andre, Townsend Kristy L
School of Biology and Ecology, University of Maine, Orono, ME, USA.
Department of Neurological Surgery, The Ohio State University, 1014 Biomedical Research Tower, 460 W. 12 Avenue, Columbus, OH, USA.
iScience. 2021 Sep 11;24(10):103127. doi: 10.1016/j.isci.2021.103127. eCollection 2021 Oct 22.
Little is known about the diversity and function of adipose tissue nerves, due in part to the inability to effectively visualize the tissue's diverse nerve subtypes and the patterns of innervation across an intact depot. The tools to image and quantify adipose tissue innervation are currently limited. Here, we present a method of tissue processing that decreases tissue thickness in the z-axis while leaving cells intact for subsequent immunostaining. This was combined with autofluorescence quenching techniques to permit intact whole tissues to be mounted on slides and imaged by confocal microscopy, with a complementary means to perform whole tissue neurite density quantification after capture of tiled z-stack images. Additionally, we demonstrate how to visualize nerve terminals (the neuro-adipose nexus) in intact blocks of adipose tissue without z-depth reduction. We have included examples of data demonstrating nerve subtypes, neurovascular interactions, label-free imaging of collagen, and nerve bundle digital cross-sections.
人们对脂肪组织神经的多样性和功能了解甚少，部分原因是无法有效可视化该组织的多种神经亚型以及完整脂肪库中的神经支配模式。目前，用于成像和量化脂肪组织神经支配的工具有限。在此，我们提出一种组织处理方法，该方法可在z轴上减小组织厚度，同时使细胞保持完整以便后续进行免疫染色。这与自发荧光淬灭技术相结合，使完整的全组织能够安装在载玻片上并通过共聚焦显微镜成像，同时在捕获平铺的z轴堆叠图像后，还提供了一种对全组织神经突密度进行量化的补充方法。此外，我们展示了如何在不减小z轴深度的情况下可视化完整脂肪组织块中的神经末梢（神经 - 脂肪连接）。我们提供了一些数据示例，展示了神经亚型、神经血管相互作用、胶原蛋白的无标记成像以及神经束数字横截面。
