The following protocol represents over 10 years of experience from our lab in culturing, recording from and stimulating neuronal networks on micro-electrode arrays. Each step has been optimized so as to lead to the development of long surviving healthy neuronal networks. References are provided where available while some optimal settings we determined empirically. Prior to initiating the protocol, obtain equipment and supplies and prepare solutions as listed in the section entitled 'Materials.' The section on 'Brain Dissection' will be briefly discussed, but not demonstrated in the accompanying video as other Journal of Visualized Experiment articles cover this topic11.
