首页> 外文会议>Multiphoton Microscopy in the Biomedical Sciences VI; Progress in Biomedical Optics and Imaging; vol.7 no.12 >Structural changes at the cellular and sub-cellular level in the cerebral cortex of mice visualized by means of trans-cranial multi photon in-vivo microscopy
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Structural changes at the cellular and sub-cellular level in the cerebral cortex of mice visualized by means of trans-cranial multi photon in-vivo microscopy

机译:通过颅内多光子体内显微镜观察到的小鼠大脑皮质细胞和亚细胞水平的结构变化

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摘要

Neural cell structural change is associated with numerous processes in the normal and diseased brain. We report about a multi photon laser scanning microscope setup that permits visualization of such neuronal and glial structural dynamics in living mice and recent observations performed by means of this instrument. The modular system consists of a modified industrial standard CSLM and is to a large degree composed of commercially available components. From a technical point of view, two developments are essential: Firstly, a multifunctional stage adapted to both laser scanning microscopic observation and pre-observational surgery has been designed and built. Secondly, the essential component of the detection unit is a state-of-the-art photomultiplier tube installed in a Peltier cooled thermal box shielding the detector from both room temperature and distortions caused by external electromagnetic fields. An electro-optical modulator is used to accurately adjust the power of the applied laser beam and to blank the beam when no data are sampled, e.g. in the dead time intervals between adjacent pixels. Photo bleaching and thermal damage are thus kept on a minimum level. In transgenic mice expressing fluorescent proteins in neuronal or glial cells, details such as spines and astrocytic endfeet can be imaged trans-cranially to at least 80 microns below the brain surface. Individual cells and cell compartments can be visualized repeatedly over extensive periods of time.
机译:神经细胞结构变化与正常和患病大脑中的许多过程有关。我们报告了一种多光子激光扫描显微镜的设置,该设置允许在活体小鼠中可视化这种神经元和神经胶质结构动力学,以及借助该仪器进行的近期观察。模块化系统由修改后的工业标准CSLM组成,并且很大程度上由市售组件组成。从技术的角度来看,两个发展是必不可少的:首先,已设计并建造了一个适用于激光扫描显微镜观察和观察前手术的多功能平台。其次,检测单元的基本组成部分是安装在珀尔帖冷却热箱中的先进光电倍增管,可保护检测器免受室温和外部电磁场引起的变形的影响。电光调制器用于精确地调节所施加的激光束的功率,并在没有数据采样时,例如在没有数据采样时将其消隐。在相邻像素之间的空载时间间隔中。因此,将光漂白和热损伤保持在最低水平。在神经元或神经胶质细胞中表达荧光蛋白的转基因小鼠中,诸如刺和星形胶质尾脚的细节可以通过颅底成像到大脑表面以下至少80微米。单个细胞和细胞室可以在很长一段时间内重复显示。

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