In Vivo Imaging of Myelin in the Vertebrate Central Nervous System Using Third Harmonic Generation Microscopy
In Vivo Imaging of Myelin in the Vertebrate Central Nervous System Using Third Harmonic Generation Microscopy
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DOI：
10.1016/j.bpj.2011.01.031
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发表时间：
2011-03-02
影响因子：
3.4
通讯作者：
Schaffer, Chris B.
中科院分区：
文献类型：
--
作者：
Farrar, Matthew J.;Wise, Frank W.;Schaffer, Chris B.
中枢神经系统（CNS）中髓鞘的损失导致衰弱性神经功能缺损。动物模型中枢神经系统中髓鞘的高分辨率光学成像受到缺乏体内髓鞘标记策略的限制。我们证明了三次谐波发生（THG）显微镜-一种相干的，非线性的，无染料的成像方式提供了小鼠中枢神经系统中髓鞘的微米分辨率成像。在固定的组织中，我们发现THG信号来自白色物质束，并与来自髓鞘特异性染料的双光子激发荧光（2 PEF）共定位。在体内，我们同时使用THG和2 PEF成像的小鼠脊髓，以解决髓鞘周围的个别荧光标记的轴突，脊髓损伤后髓鞘破坏。最后，我们提出了THG的髓鞘特异性的光学机制。这些结果确立了THG显微镜作为研究髓鞘丢失和恢复的理想工具。
Loss of myelin in the central nervous system (CNS) leads to debilitating neurological deficits. High-resolution optical imaging of myelin in the CNS of animal models is limited by a lack of in vivo myelin labeling strategies. We demonstrated that third harmonic generation (THG) microscopy-a coherent, nonlinear, dye-free imaging modality provides micrometer resolution imaging of myelin in the mouse CNS. In fixed tissue, we found that THG signals arose from white matter tracts and were colocalized with two-photon excited fluorescence (2PEF) from a myelin-specific dye. In vivo, we used simultaneous THG and 2PEF imaging of the mouse spinal cord to resolve myelin sheaths surrounding individual fluorescently-labeled axons, and followed myelin disruption after spinal cord injury. Finally, we suggest optical mechanisms that underlie the myelin specificity of THG. These results establish THG microscopy as an ideal tool for the study of myelin loss and recovery.