Brazilian Biosciences National Laboratory (LNBio), Brazilian Center for Research in Energy and Materials (CNPEM), Zip Code 13083-970, Campinas, Sao Paulo, Brazil.
Brazilian Synchrotron Light National Laboratory (LNLS), Brazilian Center for Research in Energy and Materials (CNPEM), Zip Code 13083-970, Campinas, Sao Paulo, Brazil.
Sci Rep. 2018 Aug 13;8(1):12074. doi: 10.1038/s41598-018-30501-x.
The assessment of neuronal number, spatial organization and connectivity is fundamental for a complete understanding of brain function. However, the evaluation of the three-dimensional (3D) brain cytoarchitecture at cellular resolution persists as a great challenge in the field of neuroscience. In this context, X-ray microtomography has shown to be a valuable non-destructive tool for imaging a broad range of samples, from dense materials to soft biological specimens, arisen as a new method for deciphering the cytoarchitecture and connectivity of the brain. In this work we present a method for imaging whole neurons in the brain, combining synchrotron-based X-ray microtomography with the Golgi-Cox mercury-based impregnation protocol. In contrast to optical 3D techniques, the approach shown here does neither require tissue slicing or clearing, and allows the investigation of several cells within a 3D region of the brain.
评估神经元数量、空间组织和连接对于全面理解大脑功能至关重要。然而，在神经科学领域，以细胞分辨率评估三维（3D）大脑细胞结构仍然是一个巨大的挑战。在这种情况下，X 射线显微断层扫描已被证明是一种非常有价值的非破坏性工具，可用于成像从高密度材料到软生物标本等广泛的样本，成为破译大脑细胞结构和连接的新方法。在这项工作中，我们提出了一种在大脑中成像全神经元的方法，将基于同步加速器的 X 射线显微断层扫描与戈尔吉-考克斯基于汞的浸渍方案相结合。与光学 3D 技术相比，这里展示的方法既不需要组织切片或清除，也允许在大脑的 3D 区域内研究多个细胞。
