Recent in vitro electrophysiological studies have revealed that neighboring interneurons interact with each other in a sub-millisecond time range via gap junctions and that individual dendritic compartments generate local excitation spikes and back-propagated spikes within a single-neuron. However, most in vivo electrophysiological studies using behaving animals only focus on activity rates of single-neurons and/or large neuronal populations without considering the potential role of such sub-millisecond interactions among neurons. This neglect is due to the limitation of ordinary in vivo multi-neuronal recording and spike sorting techniques applied to behaving animals. Though independent component analysis (ICA) is a powerful method to overcome certain limitations, ICA has a serious problem in that the number of single-electrodes (microwires) must be more than the number of single-neurons to be recorded. Our recently-developed method has solved this limitation of ICA, but a few problems have remained: the computational load is heavy, the method can be used only for off-line, not real-time, processing, and the electrode-neuron drift problem remains unsolved. In this paper, solving all these problems, we introduce a novel system consisting of automatic and real-time spike sorting with ICA in combination with a newly developed multi-electrode, dodecatrode. The system has the potential to answer some important neurobiological questions that have not been explored in in vivo electrophysiological experiments: how sub-millisecond interactions between closely neighboring single-neurons act in freely behaving animals. The system promises to be a bridge connecting electrophysiological studies in vitro and in vivo.