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光纤式在体荧光显微成像系统在动态观测活体动物脑内神经元中的应用

石莹, 陈露岚, 姜民^*

复旦大学脑科学研究院，医学神经生物学国家重点实验室，上海 200032

摘要

本文旨在利用光纤式在体荧光显微成像系统，建立大鼠脑内神经元的在体显微荧光连续观察的实验方法。将含有绿色荧光蛋白的重组病毒载体(Ubi-GFP)微注射到Sprague Dawley (SD)大鼠大脑皮层区，7天后用微创手术将光纤探头植入动物脑内目标位置，使用在体荧光显微成像技术观察到微注射区GFP标记神经元的特异性荧光信号。随后对大鼠脑组织行冰冻切片，荧光显微镜对切片的观察结果验证了在体荧光显微成像实验中观察到的荧光信号。该方法既保证了在体实验中动物处于生理状态，又满足了体内神经组织荧光显微水平成像的要求，可以用于动物脑内神经元荧光信号的在体追踪记录，为在体神经科学实验提供了有效的技术保障。

关键词： 在体荧光成像技术; 绿色荧光蛋白; 神经元

分类号：Q7

[An application of the fibered fluorescence microscopy to continuously monitor the rat cerebral neurons in vivo.] [Article in Chinese]

SHI Ying, CHEN Lu-Lan, JIANG Min^*

Institutes of Brain Science and State Key Laboratory of Medical Neurobiology, Fudan University, Shanghai 200032, China

Abstract

The aim of the present study was to establish an approach to continuously record fluorescent signals of rat cerebral cortical neurons in vivo, using the novel system composed of fiber-optic probe and fluorescence microscopy. To visualize cortical neurons, recombinant virus vectors carrying green fluorescent protein (GFP) gene were microinjected into cerebral cortex in Sprague Dawley (SD) rats. Seven days later, imaging microprobe, composed of optical minifibers, was inserted into the microinjected region of cerebral cortex. By using the fibered fluorescence microscopy, we observed fluorescent signals of cortical neurons transfected with GFP in living animals. In the brain slices from the microinjected region, the fluorescence signals of GFP were recorded using fluorescence microscopy, which confirmed the observation of the fibered fluorescence microscopy. The novel technology established in the present study maintains physical condition of experimental animal, and meets the demands of fluorescence micro-imaging in neural tissue in vivo. Application of this technology allows a direct and rapid approach tracing fluorescent signals of neurons in living animals.

Key words: in vivo molecular imaging technology; green fluorescent protein; neurons

收稿日期：2012-03-12　　录用日期：2012-03-21

通讯作者：姜民　　E-mail: minjiang@fudan.edu.cn

引用本文：

石莹, 陈露岚, 姜民. 光纤式在体荧光显微成像系统在动态观测活体动物脑内神经元中的应用[J]. 生理学报 2012; 64 (6): 695-699.

SHI Ying, CHEN Lu-Lan, JIANG Min. [An application of the fibered fluorescence microscopy to continuously monitor the rat cerebral neurons in vivo.] [Article in Chinese]. Acta Physiol Sin 2012; 64 (6): 695-699 (in Chinese with English abstract).