Caenorhabditis elegans (C. elegans) is an excellent model organism for the study of aging and longevity. In this work, we presented a microfluidic approach for the evaluation of longevity in C. elegans under stress. The microfluidic device integrated multiple microvalves with parallel channels, which enabled the long-term culture and flexible manipulation of C. elegans in real-time. The utility of the device was demonstrated by characterizing the lifespan, mobility behavior and fluorescence expression of oxidative stress in mutant strain CL2166 simultaneously at single animal resolution. A certain dose of polydatin was found to enable the extension of mean lifespan of CL2166 for the first time, and the prolonged longevity activity was possibly mediated by the protective response to oxidative stress, indicating the promising role of polydatin involved in aging process. The device is simple to operate, easy for real-time imaging and multiparatemer evaluations in parallel, providing the powerful platform for drug evaluation/screening in highthroughput format.