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JoVE Encyclopedia of Experiments
Neuroscience
EAE 小鼠模型中的生物发光成像
EAE 小鼠模型中的生物发光成像
Encyclopedia of Experiments
Neuroscience
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Encyclopedia of Experiments Neuroscience
Bioluminescence Imaging in an EAE Mouse Model

EAE 小鼠模型中的生物发光成像

Protocol
421 Views
02:27 min
July 8, 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

从患有实验性自身免疫性脑脊髓炎或EAE的麻醉小鼠开始。这种情况会导致眼睛和大脑发炎。

将

鼠标放入成像设备中,将上背部对齐在中心。

用黑布盖住鼠标的下部,以防止该区域的信号干扰。

使用成像软件记录鼠标位置。

取装有含有生物发光底物的试剂的注射器,腹腔注射到小鼠体内。

试剂循环并进入炎症部位,底物与炎症特异性酶相互作用。

酶-底物反应产生生物发光信号。

定期

捕获图像以监测该信号。

视神经位于表面附近,使眼睛周围的生物发光突出。相比之下,当头骨吸收发射的光时,来自大脑的信号是微弱的。

在

图像中,蓝色表示轻度炎症,红色表示严重炎症。

要进行体内成像,请在所有成像过程中将小鼠保持在2至2.5%的异氟醚麻醉下。将两只小鼠并排放置在设备中,并将小鼠的上棘置于中心。

接下来,用黑布遮盖免疫部位。打开图像捕获软件并初始化相机。然后,拍摄基线图像以评估小鼠的正确定位。

随后,使用与相机保持 6.5 厘米距离的 B 焦距来捕捉所有图像。然后,在填充注射器之前,将即用型化学发光试剂以每毫升 40 毫克混合。然后,腹膜内注射100微升小鼠,并在注射后5,10和15分钟捕获生物发光图像。

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