The emergence of new drugs is often driven by the escalating resistance of parasites to existing drugs and the accessibility of more advanced technology platforms. Microfluidic devices have allowed for quicker testing of compounds, regulated sampling/sorting of entire animals, and automated behavioral pattern detection. In the majority of microfluidic devices, the effects of drugs on small animals (e.g. Caenorhabditis elegans)elegant are quantified by an endpoint, dose response curve that shows one parameter (such as worm velocity or stroke frequency). To expand on this, we present a multi-parameter extraction method to examine the modes of paralysis in C. elegans over an extended period of time. A microfluidic device that includes real-time imaging was utilized to expose C. elegans to four anthelmintic drugs (i.e., pyrantel, levamisole, tribendimidine, and methyridine). We measured worm behavior with parameters such as curls per second, the types of paralysis, the frequency of modes, and the number/duration of active/immobilization periods. Each drug was used at EC75, where 75% of the worm population responded to the drug. While using the same concentrations, we noticed disparities in the way that worms paralyzed in the drug-filled environments. Our study stresses the importance of assessing drug effects on small animals with multiple parameters calculated at regular intervals over a prolonged time span in order to accurately capture the resistance and adaptability in chemical environments.
