Abstract

Embracing Chaos – A Simplified Platform for Multiplexing Digital Assays in Polydisperse Droplets

Samantha Byrnes, Research Scientist, Intellectual Ventures Laboratory

The development of droplet-based assays enables highly-sensitive biomarker detection and quantification, but the required equipment is often expensive, cumbersome, and challenging to operate. These constraints have isolated this technology to centralized laboratories requiring highly trained technicians. To overcome these limitations and make this technology applicable for a variety of settings, we have developed a simplified platform for performing multiplexed digital assays. Our platform reduces the complexity of both droplet creation and detection by removing the constraint of monodisperse droplets. We have developed a statistical framework to apply to assays performed in polydisperse droplets prepared without any specialized equipment. Our assays are quantified using a simplified reader developed in our lab that does not rely on highly specialized microfluidics to control detection by droplet size. Also, our current reader is designed to detect up to a 5-plexed assay. We have also developed multiple assay types that can be performed with our simplified digital platform including rapid bacterial culture, dPCR, and digital immunoassays. Additionally, we have developed a new method to increase the multiplexing capabilities of our digital system up to a 10-plex assay. Our systems shows comparable results to commercial digital systems such as BioRad’s ddPCR machine and Quanterix’s Simoa technology.