A miniature head-mounted two-photon microscope: High-resolution brain imaging in freely moving animals
A miniature head-mounted two-photon microscope: High-resolution brain imaging in freely moving animals
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DOI：
10.1016/s0896-6273(01)00421-4
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发表时间：
2001-09-27
期刊：
影响因子：
16.2
通讯作者：
Denk, W
中科院分区：
文献类型：
--
作者：
Helmchen, F;Fee, MS;Denk, W
双光子显微镜已经能够在大鼠的完整脑中进行解剖和功能荧光成像。在这里，我们通过使用小型化的头戴式显微镜将双光子成像从麻醉的、头部稳定的动物扩展到清醒的、自由移动的动物。激发光在单模光纤中传导到显微镜，并且使用光纤尖端的振动来扫描图像。显微镜性能的特点首先在麻醉大鼠的新皮层。我们很容易获得充满荧光标记血液的血管系统和充满钙指示剂的2/3层锥体神经元的图像。毛细血管血流量和树突状钙瞬变测量高时间分辨率使用线扫描。在清醒，自由移动的大鼠，稳定的成像是可能的，除非在突然的头部运动。
Two-photon microscopy has enabled anatomical and functional fluorescence imaging in the intact brain of rats. Here, we extend two-photon imaging from anesthetized, head-stabilized to awake, freely moving animals by using a miniaturized head-mounted microscope. Excitation light is conducted to the microscope in a single-mode optical fiber, and images are scanned using vibrations of the fiber tip. Microscope performance was first characterized in the neocortex of anesthetized rats. We readily obtained images of vasculature filled with fluorescently labeled blood and of layer 2/3 pyramidal neurons filled with a calcium indicator. Capillary blood flow and dendritic calcium transients were measured with high time resolution using line scans. In awake, freely moving rats, stable imaging was possible except during sudden head movements.