Activity-dependent myelination modulates neuron conduction velocity and as such it is essential for a correct wiring of a whole nervous system. Increasing myelination through inducing neuron activity has been proposed as a treatment strategy for demyelination diseases. Yet, the mechanisms and the effects of activity-dependent myelination remain elusive—new tools are needed. In this chapter, we describe a novel compartmentalized device integrated with an optogenetic stimulator for studying activity-dependent myelination in vitro. The platform can be modified to include multiple cell types, stimulation modes, and experimental readouts to answer a specific research question. This versatility combined with a precise control over spatial extent of the stimulation and the stimulation pattern make the proposed platform a valuable tool for molecular myelination studies.