Live animal myelin histomorphometry of the spinal cord with video-rate multimodal nonlinear microendoscopy
Live animal myelin histomorphometry of the spinal cord with video-rate multimodal nonlinear microendoscopy
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DOI：
10.1117/1.jbo.17.2.021107
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发表时间：
2012-02-01
影响因子：
3.5
通讯作者：
Cote, Daniel
中科院分区：
文献类型：
--
作者：
Belanger, Erik;Crepeau, Joel;Cote, Daniel
细胞动力学的体内成像可以显著地使能理解神经系统疾病的病理生理学。为了充分利用这种方法的力量，主要的挑战是最大限度地减少侵入性，并最大限度地增加可以组合以探测多个细胞过程之间相互作用的并发光学信号的数量。无标记相干反斯托克斯拉曼散射（汽车）显微镜，例如，可用于跟踪神经退行性疾病或创伤后的脱髓鞘，但髓鞘成像本身不足以理解导致白色物质中出现病变的复杂事件序列。一个市售的显微内窥镜在这里使用，以实现微创，视频速率多模态非线性成像的细胞过程中活的小鼠脊髓。该系统允许同时汽车髓鞘成像和小胶质细胞和轴突的双光子激发荧光显微内窥镜。还描述了在高空间分辨率下的形态测量数据提取，具有用于减少运动相关的成像伪影的技术。尽管其直径小，显微内窥镜能够在广泛的组织区域上进行高速多模态成像，但分辨率足以量化髓鞘厚度和小胶质细胞运动性的细微差异。(C)2012年，美国光电仪器工程师学会（SPIE）。[DOI：10.1117/1.JBO.17.2.021107]
In vivo imaging of cellular dynamics can be dramatically enabling to understand the pathophysiology of nervous system diseases. To fully exploit the power of this approach, the main challenges have been to minimize invasiveness and maximize the number of concurrent optical signals that can be combined to probe the interplay between multiple cellular processes. Label-free coherent anti-Stokes Raman scattering (CARS) microscopy, for example, can be used to follow demyelination in neurodegenerative diseases or after trauma, but myelin imaging alone is not sufficient to understand the complex sequence of events that leads to the appearance of lesions in the white matter. A commercially available microendoscope is used here to achieve minimally invasive, video-rate multimodal nonlinear imaging of cellular processes in live mouse spinal cord. The system allows for simultaneous CARS imaging of myelin sheaths and two-photon excitation fluorescence microendoscopy of microglial cells and axons. Morphometric data extraction at high spatial resolution is also described, with a technique for reducing motion-related imaging artifacts. Despite its small diameter, the microendoscope enables high speed multimodal imaging over wide areas of tissue, yet at resolution sufficient to quantify subtle differences in myelin thickness and microglial motility. (C) 2012 Society of Photo-Optical Instrumentation Engineers (SPIE). [DOI: 10.1117/1.JBO.17.2.021107]