Neurotransmitters have been well known as information carriers for a long time. Recently, some of the research indicated their neurotoxicity, while some indicated their neurotrophic actions. It is very important to understand the role of neurotransmitters. Glutamate is one of the most important excitatory neurotransmitter in the brain. We developed a novel measurement method for glutamate. The method we describe here is based on the enzyme-mediated electrochemical detection. Glutamate oxidase and horseradish peroxidase were deposited together with polymer-mediator on the electrode. We applied this idea on ITO multi-array electrode and developed a 64 channel multi-array sensor. The sensor permits us to detect glutamate release from multiple regions simultaneously in real time. As it is possible to illustrate the distribution of glutamate release, the sensor could be used not only in the pharmacological field, but also in medical treatment in the near future.