Interfaces provide the structural basis of essential bone functions. In the hierarchical structure of bone tissue, heterogeneities such as porosity or boundaries are found at scales ranging from nanometers to millimeters, all of which contributing to macroscopic properties. To date, however, the complexity or limitations of currently used imaging methods restrict our understanding of this functional integration. Here we address this issue using label-free third-harmonic generation (THG) microscopy. We find that the porous lacuno-canalicular network (LCN), revealing the geometry of osteocytes in the bone matrix, can be directly visualized in 3D with submicron precision over millimetric fields of view compatible with histology. THG also reveals interfaces delineating volumes formed at successive remodeling stages. Finally, we show that the structure of the LCN can be analyzed in relation with that of the extracellular matrix and larger-scale structures by simultaneously recording THG and second-harmonic generation (SHG) signals relating to the collagen organization.

译文

:接口提供基本的骨骼功能的结构基础。在骨组织的分层结构中,发现异质性(例如孔隙度或边界)的范围从纳米到毫米,所有这些都有助于宏观特性。然而,迄今为止,当前使用的成像方法的复杂性或局限性限制了我们对这种功能集成的理解。在这里,我们使用无标记的三次谐波(THG)显微镜来解决此问题。我们发现,多孔的腔管网络(LCN)揭示了骨基质中骨细胞的几何形状,可以在与组织学兼容的毫米视野中以亚微米精度直接以3D可视化。 THG还揭示了描述在连续重塑阶段形成的体积的界面。最后,我们表明,通过同时记录与胶原组织有关的THG和第二谐波产生(SHG)信号,可以分析LCN的结构与细胞外基质的结构和更大尺度的结构。

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