Kogan A, Ross W N, Zecevic D, Lasser-Ross N
Department of Neurobiology, Hebrew University, Givat-Ram, Jerusalem, Israel.
Brain Res. 1995 Nov 27;700(1-2):235-9. doi: 10.1016/0006-8993(95)00956-q.
We evaluated several techniques for their ability to record membrane potential changes with voltage-sensitive dyes introduced into CNS neurons in the brain slice preparation. Using a probe designed for intracellular application, JPW1114, we found that iontophoresis or pressure pulses could not push the lipophilic dye through electrodes whose resistance was sufficiently high to produce good electrical recordings in cerebellar Purkinje neurons. However, properly selected patch electrodes could introduce the dye into the cell and still give good electrical records. Using this technique we recorded depolarizing and hyperpolarizing transients and climbing fiber responses using either a single photodiode or a fast, cooled CCD camera. While these results are promising, there are still problems due to the slow diffusion of the dye in the dendrites and a low sensitivity which requires signal averaging to acquire traces with a good signal to noise ratio.
我们评估了几种技术，以研究它们在脑片制备中，将电压敏感染料引入中枢神经系统神经元来记录膜电位变化的能力。使用一种专为细胞内应用设计的探针JPW1114，我们发现离子电渗法或压力脉冲无法将亲脂性染料推送通过电阻足够高、能在小脑浦肯野神经元中产生良好电记录的电极。然而，正确选择的膜片电极可以将染料引入细胞，同时仍能给出良好的电记录。使用该技术，我们通过单个光电二极管或快速冷却的电荷耦合器件相机记录了去极化和超极化瞬变以及攀缘纤维反应。虽然这些结果很有前景，但由于染料在树突中的扩散缓慢以及灵敏度较低，仍存在问题，这需要进行信号平均以获取具有良好信噪比的记录轨迹。
