<p>Microfluidic devices, including lab-on-a-chip devices, have many advantages over conventional laboratory techniques. Although common in some applications, there are several barriers to wider adoption, including the high initial cost to fabricate a design and the labor-intensive process of development. This work seeks to address those barriers by combining 3D printing and computer aided design tools. Building on existing open-source software from electronic design automation (EDA), we present a design, verification, and manufacturing toolchain for 3D design for microfluidic devices. The process starts with a list of components and connections then, automatically lays out a microfluidic device using a library of components, simulates the device, and produces a 3D CAD file that is used for DLP 3D printing process. The automated design and fabrication process was demonstrated by automatically designing and fabricating a calcium quantification assay. This toolchain automatically generated a microfluidic chip that meters each reagent with an error of less than 2.24% as verified by Xyce simulation of the chip. Physical chips were printed and found to perform with errors less than 9.2% on average compared to the assay performed by hand. The demonstration showed the ability of the toolchain to automatically generate a functional microfluidic chip for use with real assays using previously developed EDA tools.</p>