The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project will be the development of a 3-dimensional (3D) culture well plate to be used as a high-throughput screening (HTS) tool for research and drug discovery. The core technology is a novel well plate that incorporates micron-meter-sized channels and compartments, and will have the ability to perform autonomous media exchange and drug loading non-invasively in an automated and high-speed fashion. The well plate will allow larger libraries of potential drug candidate compounds to be screened against 3D cultured cells. This would enable pharmaceutical companies and screening centers to increase the probability of finding new drug candidates by elevating the scale and throughput necessary for their discovery and validation. The technology also will increase scientific understanding of cells by providing a tool to study their behavior in an in vivo-like environment, which is important when studying tumor spheroids and exploring cancer therapeutics. This SBIR Phase I project proposes to develop a 3D cell culture tool by integrating novel microfluidic technology into a well plate format. 3D cell culture is emerging as the next generation drug screening method, as it provides a window into cellular responses by culturing and studying cells in an in vivo-like environment. Currently, however, there are no tools available in the market for testing 3D cultured cells against drug compounds in an HTS fashion. A prototype of the well plate will be fabricated and several important parameters that are required to conduct HTS studies on tumors will be studied. Parameters such as the uniformity in culturing 3D tumors, volume of media required for healthy cellular growth, and effectiveness of the plate to perform screening of drug compounds will be determined. It is anticipated that the microfluidic well plate will create a uniform number of tumors in each microfluidic well and produce results similar to those derived from the current state of the art 3D cell culture platforms. Proprietary patented microfluidic technology developed and verified during the SBIR Phase I project will help integrate 3D cell culture into the mainstream drug discovery cascade.