Investigation of human multiple sclerosis lesions using high resolution spectrally unmixed CARS microscopy
Investigation of human multiple sclerosis lesions using high resolution spectrally unmixed CARS microscopy
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DOI：
10.1117/12.2005504
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发表时间：
2013-03
期刊：
影响因子：
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通讯作者：
K. Poon;C. Brideau;Wulin Teo;G. Schenk;Roel Klaver;A. Klauser;J. Kawasoe;J. Geurts;P. Stys
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文献类型：
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作者：
K. Poon;C. Brideau;Wulin Teo;G. Schenk;Roel Klaver;A. Klauser;J. Kawasoe;J. Geurts;P. Stys
多发性硬化（MS）的病理学涉及脑和脊髓的灰质和白色物质区域。其特征在于慢性病变中脱髓鞘、炎性浸润、轴突变性和随后的胶质增生的各种组合。虽然急性和慢性白色斑块的特征很好，很容易识别，但有证据表明MS患者的CNS可能发生全面改变，在外观正常的白色物质（NAWM）和弥漫性异常的白色物质（DAWM）中发现细微异常，其中组织化学染色和高级磁共振成像表明组织成分改变。因此，典型的急性炎性病变可能仅仅是中枢神经系统慢性广泛受累的最明显的表现，难以可靠地检查。目前的研究涉及的显微结构和生物化学的脱髓鞘，髓鞘再生和轴突损失在不同地区的死后人类MS脑，包括NAWM，髓鞘再生领域和更典型的急性和慢性病变。使用一种新型的相干反斯托克斯拉曼散射（汽车）显微镜与双光子激发荧光（TPEF）同步成像研究髓鞘，神经胶质细胞和血管周围空间。探测~ 2800和3000 cm-1之间的活性CH拉伸区域，以提供与疾病进展有关的化学特异性、高分辨率、无标记成像。汽车数据与TPEF和常规组织化学和免疫组织化学染色相关。我们的新型汽车显微镜系统提供了详细的形态学和生物化学信息的CNS病理MS和可能适用于广泛的其他人类大脑和脊髓疾病。
The pathology of multiple sclerosis (MS) involves both the gray and white matter regions of the brain and spinal cord. It is characterized by various combinations of demyelination, inflammatory infiltration, axonal degeneration, and later gliosis in chronic lesions. While acute and chronic white matter plaques are well characterized and easily identified, evidence indicates that the CNS of MS patients may be globally altered, with subtle abnormalities found in grossly normal appearing white matter (NAWM) and in diffusely abnormal white matter (DAWM) where histochemical stains and advanced magnetic resonance imaging indicate altered tissue composition. Thus, the prototypical acute inflammatory lesion may merely represent the most obvious manifestation of a chronic widespread involvement of the CNS, which is difficult to examine reliably. The current study deals with the microstructure and biochemistry of demyelination, remyelination and axonal loss in various regions of post-mortem human MS brain, including NAWM, areas of remyelination and more typical acute and chronic lesions. The myelin sheath, neuroglia and perivascular spaces were investigated using a novel Coherent Anti-Stokes Raman Scattering (CARS) microscope with simultaneous Two-Photon Excited Fluorescence (TPEF) imaging. The active CH stretching region between ~ 2800 and 3000 cm-1 was probed to provide chemically specific, high resolution, label-free imaging pertaining to the progression of the disease. CARS data were correlated with TPEF and conventional histochemical and immunohistochemical stains. Our novel CARS microscopy system provides detailed morphological and biochemical information regarding CNS pathology in MS and that may be applicable to a broad range of other human brain and spinal cord disorders.