Shi Ying, Chen Lu-Lan, Jiang Min
Institutes of Brain Science and State Key Laboratory of Medical Neurobiology, Fudan University, Shanghai 200032, China.
Sheng Li Xue Bao. 2012 Dec 25;64(6):695-9.
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.
本研究的目的是建立一种使用由光纤探头和荧光显微镜组成的新型系统在体内连续记录大鼠大脑皮质神经元荧光信号的方法。为了可视化皮质神经元,将携带绿色荧光蛋白(GFP)基因的重组病毒载体微量注射到Sprague Dawley(SD)大鼠的大脑皮质中。7天后,将由光学微纤维组成的成像微探头插入大脑皮质的微量注射区域。通过使用光纤荧光显微镜,我们在活体动物中观察到了转染GFP的皮质神经元的荧光信号。在来自微量注射区域的脑片中,使用荧光显微镜记录了GFP的荧光信号,这证实了光纤荧光显微镜的观察结果。本研究中建立的新技术保持了实验动物的生理状态,并满足了体内神经组织荧光微成像的要求。应用该技术可以直接、快速地追踪活体动物中神经元的荧光信号。