Chi Jingyi, Crane Audrey, Wu Zhuhao, Cohen Paul
Laboratory of Molecular Metabolism, The Rockefeller University.
Laboratory of Brain Development and Repair, The Rockefeller University; Laboratory of Molecular Genetics, The Rockefeller University;
J Vis Exp. 2018 Jul 28(137):58271. doi: 10.3791/58271.
Adipose tissue plays a central role in energy homeostasis and thermoregulation. It is composed of different types of adipocytes, as well as adipocyte precursors, immune cells, fibroblasts, blood vessels, and nerve projections. Although the molecular control of cell type specification and how these cells interact have been increasingly delineated, a more comprehensive understanding of these adipose-resident cells can be achieved by visualizing their distribution and architecture throughout the whole tissue. Existing immunohistochemistry and immunofluorescence approaches to analyze adipose histology rely on thin paraffin-embedded sections. However, thin sections capture only a small portion of tissue; as a result, the conclusions can be biased by what portion of tissue is analyzed. We have therefore developed an adipose tissue clearing technique, Adipo-Clear, to permit comprehensive three-dimensional visualization of molecular and cellular patterns in whole adipose tissues. Adipo-Clear was adapted from iDISCO/iDISCO+, with specific modifications made to completely remove the lipid stored in the tissue while preserving native tissue morphology. In combination with light-sheet fluorescence microscopy, we demonstrate here the use of the Adipo-Clear method to obtain high-resolution volumetric images of an entire adipose tissue.
脂肪组织在能量平衡和体温调节中起着核心作用。它由不同类型的脂肪细胞以及脂肪细胞前体、免疫细胞、成纤维细胞、血管和神经突起组成。尽管细胞类型特异性的分子控制以及这些细胞如何相互作用已越来越清晰，但通过可视化它们在整个组织中的分布和结构，可以更全面地了解这些驻留在脂肪组织中的细胞。现有的用于分析脂肪组织学的免疫组织化学和免疫荧光方法依赖于薄的石蜡包埋切片。然而，薄切片仅捕获组织的一小部分；因此，结论可能会因所分析的组织部分而产生偏差。因此，我们开发了一种脂肪组织透明化技术——Adipo-Clear，以实现对整个脂肪组织中分子和细胞模式的全面三维可视化。Adipo-Clear是在iDISCO/iDISCO+的基础上改进而来，进行了特定的修改以完全去除组织中储存的脂质，同时保留天然组织形态。结合光片荧光显微镜，我们在此展示了使用Adipo-Clear方法获得整个脂肪组织的高分辨率体积图像。
