Autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains.
Autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains.
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DOI：
10.1038/s41598-021-86092-7
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发表时间：
2021-04-13
影响因子：
4.6
通讯作者：
Pavone FS
中科院分区：
文献类型：
--
作者：
Costantini I;Baria E;Sorelli M;Matuschke F;Giardini F;Menzel M;Mazzamuto G;Silvestri L;Cicchi R;Amunts K;Axer M;Pavone FS
在连接组学中，用大体积的单轴突分辨率分析神经纤维的结构是一个挑战。不同的技术试图解决这一目标;然而，它们受到纤维的无效标记或可实现的分辨率的限制。区分不同相邻有髓轴突的可能性提供了提供更多关于特定区域内的纤维组成和结构的信息的机会。在这里，我们提出了MAGIC（髓鞘自发荧光成像甘油诱导对比增强），组织制备方法进行无标记的荧光成像的有髓纤维，是用户友好，易于处理。我们利用双光子荧光显微镜（TPFM）光学切片的高轴向和径向分辨率来破译感兴趣的样品内的各种纤维取向的混合物。我们证明了其广泛的适用性，通过执行介观重建在亚微米分辨率的小鼠，大鼠，猴子和人脑样品，并通过量化不同的纤维组织在控制和Reeler小鼠的海马部分。我们的研究提供了一种新的方法，用于在高分辨率下对固定样品中的神经纤维进行3D无标记成像，低于微米水平，克服了与有髓鞘轴突外源性标记相关的限制，提高了分析大脑连接的可能性。
Analyzing the structure of neuronal fibers with single axon resolution in large volumes is a challenge in connectomics. Different technologies try to address this goal; however, they are limited either by the ineffective labeling of the fibers or in the achievable resolution. The possibility of discriminating between different adjacent myelinated axons gives the opportunity of providing more information about the fiber composition and architecture within a specific area. Here, we propose MAGIC (Myelin Autofluorescence imaging by Glycerol Induced Contrast enhancement), a tissue preparation method to perform label-free fluorescence imaging of myelinated fibers that is user friendly and easy to handle. We exploit the high axial and radial resolution of two-photon fluorescence microscopy (TPFM) optical sectioning to decipher the mixture of various fiber orientations within the sample of interest. We demonstrate its broad applicability by performing mesoscopic reconstruction at a sub-micron resolution of mouse, rat, monkey, and human brain samples and by quantifying the different fiber organization in control and Reeler mouse's hippocampal sections. Our study provides a novel method for 3D label-free imaging of nerve fibers in fixed samples at high resolution, below micrometer level, that overcomes the limitation related to the myelinated axons exogenous labeling, improving the possibility of analyzing brain connectivity.
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影响因子：
3
作者：
Heath F;Hurley SA;Johansen-Berg H;Sampaio-Baptista C
通讯作者：
Sampaio-Baptista C
微光学断层扫描获得小鼠大脑的高分辨率图谱
影响因子：
56.9
作者：
Li, Anan;Gong, Hui;Luo, Qingming
通讯作者：
Luo, Qingming
影响因子：
2.4
作者：
Marchetti, Marco;Baria, Enrico;Pavone, Francesco Saverio
通讯作者：
Pavone, Francesco Saverio
影响因子：
2.9
作者：
LEDEEN, RW;HALEY, JE
通讯作者：
HALEY, JE
影响因子：
4.3
作者：
Franceschini A;Costantini I;Pavone FS;Silvestri L
通讯作者：
Silvestri L