<p>Brain tissue orchestrates neuronal function through biochemical and mechanical cues. Utilizing in vitro modeling, often the dynamics of mechanical aspects in neuronal cell cultures is neglected. However, the growing recognition of the importance of mechanical cues in neural development and healthy brain function necessitates a shift in how we study cultured neurons. Microfluidic platforms, like a Brain-on-Chip (BoC), can take active mechanical stimuli into account. In our BoC design a set of microchannels manufactured in a glass substrate by FEMTOprint technology is assembled with a spin-coated polydimethylsiloxane (PDMS) membrane and a PDMS culture chamber, which was fabricated from a stereolithographically made mold by replication. The membrane can locally deform across the culture chamber by air pressure. This paper describes the design, fabrication and test of such a novel BoC, offering an experimental setting in which we demonstrated mechano-dynamic elevated Calcium signaling in cultured human induced neural stem cell-derived neuronal networks.</p>