Significance Advancing our understanding of the underlying molecular mechanisms of aging, as well as their contributions to age-associated diseases, will have a profound impact on public health. Studying the replicative aging phenomenon in the budding yeast Saccharomyces cerevisiae has led to significant findings on how aging is regulated by evolutionarily conserved enzymes and molecular pathways. We have developed a microfluidic system that enables the visualization and analysis of the complete replicative lifespan of a single yeast cell. This system overcomes current technical challenges in low-throughput yeast lifespan analysis by providing a fast, high-throughput, and accurate analytical method at the single-cell level. This approach opens a new avenue for aging and longevity research using yeast genetic screens.