Köhling R, Greiner C, Wölfer J, Wassmann H, Speckmann E J
Institut für Physiologie, Westfälische Wilhelms-Universität, Münster, Germany.
J Neurosci Methods. 1998 Dec 1;85(2):181-6. doi: 10.1016/s0165-0270(98)00138-1.
For investigations of hypoxic effects in nervous tissue, brain slices are often used as a model system. This provides the advantage that parameters of the micromilieu, e.g. pH and temperature can easily be controlled and measurements of different data, e.g. bioelectric potentials, ion activities etc. can be performed. It is of special importance that the PO2 the slice preparation is exposed to is equally controlled under these conditions. Therefore, a PO2 monitoring system is needed which provides representative values for the tissue environment. This requirement is fulfilled by an optical PO2 sensing method based on phosphorescence quenching as a function of PO2. Here, the application of this method as adapted for use in in vitro models is described and compared to the polarographic oxygen-sensing method. Both the optical and polarographic methods are comparable regarding accuracy and response time of measurements. Furthermore, both the optical method and electrophysiological measurements can be combined. Lastly, under experimental conditions, neither the phosphorescent dye Palladium-meso-tetra-4-carboxyphenyl-porphine nor the illumination necessary for excitation of the dye influence bioelectric activity of neuronal tissue in vitro. In conclusion, the optical PO2 sensing method presented here provides a tool for reliable and continuous monitoring of PO2 in the immediate environment of brain slice preparations.
在对神经组织中的缺氧效应进行研究时，脑片常被用作模型系统。这样做的优点是，微环境参数，如pH值和温度能够轻松得到控制，并且可以对不同数据进行测量，如生物电位、离子活性等。在这些条件下，特别重要的是要对脑片制备物所暴露的PO2进行同等控制。因此，需要一个PO2监测系统，它能为组织环境提供具有代表性的值。基于磷光猝灭与PO2的函数关系的光学PO2传感方法满足了这一要求。在此，将描述这种适用于体外模型的方法的应用，并与极谱氧传感方法进行比较。在测量的准确性和响应时间方面，光学方法和极谱方法具有可比性。此外，光学方法和电生理测量都可以结合起来。最后，在实验条件下，磷光染料钯-中-四-4-羧基苯基卟啉以及激发该染料所需的光照都不会影响体外神经元组织的生物电活性。总之，本文介绍的光学PO2传感方法为可靠且连续地监测脑片制备物周围环境中的PO2提供了一种工具。
