Microfluidic systems play a crucial role in brain-on-chip research due to the precise control of fluid delivery at much smaller scales. Systems like these are necessary to maintain the conditions needed for the tissue’s health and to regulate experimental conditions. A previously in-house-developed brain-on-chip instrument relies on alternating droplets of media and oxygen to keep the tissue alive. However, this instrument presents a few limitations. First, the droplet are much more viscous than the bubble, meaning they want to adhere to each other. Due to this issue the liquid that is left behind holds a lot of cellular waste. This is harmful for the tissue and its health. Second, there is a restricted physical access to the tissue for electrophysiological measurements. The focus of this project is to design and develop a 3D printed microfluidic peristaltic pump for brain-on-chip applications. This will be produced by using commercially available tubing. This pump will be designed to provide a bidirectional fluid control using a stepper motor. Characterization will be done by analyzing effects of motor speed, tubing diameter, and synchronization of the dual channels. Pump and fluid control will be analyzed via video recording to determine optimal operating conditions. Optimization will allow for improved fluid exchange, reduced droplet retention, and allow physical access to the tissue. This project will provide a more accessible and more reproducible microfluidic peristaltic pump for brain-on-chip applications and at a low cost but efficient work rate.