Super-resolved fluorescence imaging of peripheral nerve.
Super-resolved fluorescence imaging of peripheral nerve.
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周围神经的超分辨荧光成像。
DOI：
10.1038/s41598-022-16769-0
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发表时间：
2022-07-21
影响因子：
4.6
通讯作者：
Jowett, Nate
中科院分区：
文献类型：
--
作者：
Hernandez, Ivan Coto;Mohan, Suresh;Minderler, Steven;Jowett, Nate
传统的周围神经组织病理学评价采用明视野显微镜，衍射极限分辨率约为250 nm。虽然电子显微镜可以获得神经系统的纳米级分辨率，但样品制备成本很高，而且该技术与活体样品不兼容。超分辨率显微镜（SRM）包括一组成像技术，允许使用可见光的荧光物体的纳米级分辨率。SRM的出现改变了生物医学科学，建立了无毒的手段，研究纳米细胞结构。本文中，通过超分辨率径向波动显微镜、受激发射损耗显微镜和结构化照明显微镜对来自GFP变体表达小鼠的坐骨神经切片和来自用髓鞘特异性荧光染料标记的跨面部自体移植物的再生人神经进行成像。小鼠坐骨神经轴向冷冻切片的超分辨率成像产生了强大的可视化有髓和无髓轴突。人跨面神经移植物的轴向冷冻切片的超分辨率成像显示小口径薄髓鞘再生运动轴突的分辨率增强。超分辨率成像技术提供的分辨率和对比度增强，使无髓鞘轴突，再生轴突，细胞骨架超微结构，和哺乳动物周围神经的神经元附属物使用光学显微镜的可视化。
Traditional histopathologic evaluation of peripheral nerve employs brightfield microscopy with diffraction limited resolution of ~ 250 nm. Though electron microscopy yields nanoscale resolution of the nervous system, sample preparation is costly and the technique is incompatible with living samples. Super-resolution microscopy (SRM) comprises a set of imaging techniques that permit nanoscale resolution of fluorescent objects using visible light. The advent of SRM has transformed biomedical science in establishing non-toxic means for investigation of nanoscale cellular structures. Herein, sciatic nerve sections from GFP-variant expressing mice, and regenerating human nerve from cross-facial autografts labelled with a myelin-specific fluorescent dye were imaged by super-resolution radial fluctuation microscopy, stimulated emission depletion microscopy, and structured illumination microscopy. Super-resolution imaging of axial cryosections of murine sciatic nerves yielded robust visualization myelinated and unmyelinated axons. Super-resolution imaging of axial cryosections of human cross-facial nerve grafts demonstrated enhanced resolution of small-caliber thinly-myelinated regenerating motor axons. Resolution and contrast enhancement afforded by super-resolution imaging techniques enables visualization of unmyelinated axons, regenerating axons, cytoskeleton ultrastructure, and neuronal appendages of mammalian peripheral nerves using light microscopes.
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