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JoVE Encyclopedia of Experiments
Neuroscience
使用落射荧光显微镜对膝状神经节神经元反应进行体内成像
使用落射荧光显微镜对膝状神经节神经元反应进行体内成像
Encyclopedia of Experiments
Neuroscience
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Encyclopedia of Experiments Neuroscience
In Vivo Imaging of Geniculate Ganglion Neuron Responses Using Epifluorescent Microscopy

使用落射荧光显微镜对膝状神经节神经元反应进行体内成像

Protocol
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02:13 min
August 13, 2025
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Please note that some of the translations on this page are AI generated. Click here for the English version.

Transcript

从麻醉小鼠开始,该小鼠具有手术暴露的膝状神经节,这是一组味觉处理周围神经元。

这些神经元表达 GCaMP,这是一种钙指示剂,在钙结合时发出荧光。

去除覆盖神经节的任何液体以获得清晰的可视化。

在

显微镜下将鼠标放在吸收垫上,并使用解剖标志定位膝状神经节。

在落射荧光示波器上使用适当的滤光片,检测表达GCaMP的神经元的绿色荧光。

接下来,将连接到味觉剂输送管的分配针插入小鼠的嘴中并输送味觉剂。

味觉剂刺激味觉受体细胞,释放 ATP,与神经节神经元受体结合,触发钙流入。

这些钙离子与 GCaMP 结合,诱导荧光发射。

将

相机与显微镜的视场对齐,并将视频录制与味觉传递同步,以通过荧光变化捕捉实时神经元活动。

要运行 tastant panel,请使用抽吸力从膝状体上去除液体,然后将鼠标放在解剖显微镜下的吸收垫上。使用大疱上留下的孔、颞骨上的孔和第七神经来定位膝状神经节。并在落射荧光镜上使用FITC / GFP滤光片检查单个表达GCaMP的膝状神经节神经元。将一条品尝线的分配针牢牢地放入动物的嘴里。将视频录制的开始与品尝演示的开始同步,观看实时反馈以获取响应。

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