Neuron-on-a-Chip technology is based on advanced neuronal culture technique, surface micropatterning, microelectrode array technology, and multi-dimensional data analysis techniques. The combination of these techniques allowed us to design and analyze live biological neural networks in vitro using real neurons. In this review article, two underlying technologies are reviewed: Microelectrode array technology and Neuronal patterning technology. There are new opportunities in the fusion of these technologies to apply them in neurobiology, neuroscience, neural prostheses, and cell-based biosensor areas.