Multimodal coherent anti-Stokes Raman scattering microscopy reveals microglia-associated myelin and axonal dysfunction in multiple sclerosis-like lesions in mice
Multimodal coherent anti-Stokes Raman scattering microscopy reveals microglia-associated myelin and axonal dysfunction in multiple sclerosis-like lesions in mice
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DOI：
10.1117/1.3533312
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发表时间：
2011-02-01
影响因子：
3.5
通讯作者：
Khoury, Samia J.
中科院分区：
文献类型：
--
作者：
Imitola, Jaime;Cote, Daniel;Khoury, Samia J.
髓磷脂丢失和轴突变性在许多神经系统疾病中占主导地位；然而，在活组织中同时可视化它们的方法是不可用的。我们描述了一种新的成像策略，将视频速率反射和荧光共聚焦成像与调谐到髓磷脂脂质CH2振动的相干抗斯托克斯拉曼散射（CARS）显微镜相结合，应用于慢性实验性自身免疫性脑脊髓炎（EAE）动物的活组织。我们的方法可以在高空间分辨率和信噪比的情况下监测脑切片的脱髓鞘和神经退行性变。在EAE动物的胼胝体和脊髓中可见局部严重的脂质信号缺失，表明脱髓鞘和轴突反射信号的缺失。即使在EAE小鼠的髓鞘区，髓磷脂脂质信号的强度也明显降低。在杂合敲入小鼠中，绿色荧光蛋白取代了标记中枢神经系统小胶质细胞的CX(3)CR1编码序列，我们发现激活的小胶质细胞区域与反射改变的区域和反映轴突损伤和脱髓鞘的CARS信号共定位。我们的数据证明了在多发性硬化症小鼠模型中使用多模态CARS显微镜来表征脱髓鞘和神经退行性病理，并进一步证实了小胶质细胞在慢性炎症性神经变性中的关键作用。(C) 2011光学仪器工程师学会（SPIE）(DOI: 10.1117/1.3533312)
Myelin loss and axonal degeneration predominate in many neurological disorders; however, methods to visualize them simultaneously in live tissue are unavailable. We describe a new imaging strategy combining video rate reflectance and fluorescence confocal imaging with coherent anti-Stokes Raman scattering (CARS) microscopy tuned to CH2 vibration of myelin lipids, applied in live tissue of animals with chronic experimental autoimmune encephalomyelitis (EAE). Our method allows monitoring over time of demyelination and neurodegeneration in brain slices with high spatial resolution and signal-to-noise ratio. Local areas of severe loss of lipid signal indicative of demyelination and loss of the reflectance signal from axons were seen in the corpus callosum and spinal cord of EAE animals. Even in myelinated areas of EAE mice, the intensity of myelin lipid signals is significantly reduced. Using heterozygous knock-in mice in which green fluorescent protein replaces the CX(3)CR1 coding sequence that labels central nervous system microglia, we find areas of activated microglia colocalized with areas of altered reflectance and CARS signals reflecting axonal injury and demyelination. Our data demonstrate the use of multimodal CARS microscopy for characterization of demyelinating and neurodegenerative pathology in a mouse model of multiple sclerosis, and further confirm the critical role of microglia in chronic inflammatory neurodegeneration. (C) 2011 Society of Photo-Optical Instrumentation Engineers (SPIE). [DOI:10.1117/1.3533312]