Microfluidics is the science of designing, manufacturing, and formulating processes to generate devices that are capable of analyzing small sample volumes, usually in the range of microliters (10-6) to picoliters (10-1). Microfluidic techniques have emerged as a promising alternative to conventional laboratory assays since they allow complete laboratory protocols to be performed on a single chip, merely a few square centimeters in size. Applied microfluidics have a number of significant advantages in biomedical research and in creating clinically useful technologies. For example, microfluidics enable the fabrication of new cost-effective biosensing technologies for clinical diagnostics. This cost decrease observed stems from the small scale of the device’s architecture, which requires reduced sample volumes, processing times, and reagent consumption when compared to conventional methods. At such a small scale, material selection is a crucial part of microfluidic system development as it impacts its processing, functionality, application, and the disposability of the sensor strips and the fluidic manifold. This chapter reviews the most common types of materials that are currently used to fabricate microfluidic devices. Methods used for their fabrication, physical and chemical properties of the materials, and advantages they provide to the biosensor configuration are also summarized. Special consideration was also given to the selection of ideal prototyping materials for specific applications based on their cost, mechanical, and biocompatible properties.