Objective:

Oxidative stress plays an important role in the onset of many neuronal and peripheral disorders. We examined the feasibility of obtaining semiquantitative fluorescent images of reactive oxygen species (ROS) generation in mouse brain and kidney utilizing a planar laser scanner and dihydroethidium (DHE).

Methods:

To investigate ROS generation in brain, sodium nitroprusside was injected into the striatum. Dihydroethidium was injected into the tail vein. After DHE injection, tissue slices were analyzed utilizing a planar laser scanner. For kidney study, cis-diamminedichloroplatinum [II] (cisplatin) was intraperitoneally administrated into mice.

Results:

Clear and semiquantitative fluorescent images of ROS generation in the mouse brain and kidney were obtained. Furthermore, the fluorescence intensity was stable and not affected by fading. Sodium nitroprusside induced approximately 6 times the fluorescence accumulation in the brain. Cisplatin caused renal injury in all mice, and in comparison with control mice, more than 10 times fluorescence accumulation was observed in the renal medulla with tubular necrosis and vacuolization.

Conclusions:

We successfully obtained ex vivo semiquantitative fluorescent images of ROS generation utilizing a planar laser scanner and DHE. This simple method is useful for ROS detection in several ROS-related animal models and would be applicable to a variety of biochemical processes.

中文翻译：

---

利用平面激光扫描仪进行的简单体外半定量荧光成像：检测小鼠脑和肾脏中活性氧的生成。

目的：

氧化应激在许多神经元和周围疾病的发作中起重要作用。我们检查了使用平面激光扫描仪和二氢乙锭（DHE）获得小鼠脑和肾脏中活性氧（ROS）生成的半定量荧光图像的可行性。

方法：

为了研究大脑中ROS的生成，将硝普钠注入纹状体。二氢乙锭被注入尾静脉。DHE注射后，使用平面激光扫描仪分析组织切片。为了进行肾脏研究，将腹膜内施用顺二氨二氯铂[II]（顺铂）。

结果：

获得了小鼠大脑和肾脏中ROS的清晰和半定量荧光图像。此外，荧光强度稳定并且不受褪色影响。硝普钠诱导约6倍的荧光在大脑中积累。顺铂对所有小鼠均造成肾损伤，与对照小鼠相比，在肾髓质中观察到荧光积累超过10倍，伴有肾小管坏死和空泡。

结论：

我们成功地利用平面激光扫描仪和DHE获得了ROS生成的离体半定量荧光图像。这种简单的方法可用于几种与ROS相关的动物模型中的ROS检测，并将适用于多种生化过程。