High-throughput analysis of yeast replicative aging using a microfluidic system

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. Saccharomyces cerevisiae has been an important model for studying the molecular mechanisms of aging in eukaryotic cells. However, the laborious and low-throughput methods of current yeast replicative lifespan assays limit their usefulness as a broad genetic screening platform for research on aging. We address this limitation by developing an efficient, high-throughput microfluidic single-cell analysis chip in combination with high-resolution time-lapse microscopy. This innovative design enables, to our knowledge for the first time, the determination of the yeast replicative lifespan in a high-throughput manner. Morphological and phenotypical changes during aging can also be monitored automatically with a much higher throughput than previous microfluidic designs. We demonstrate highly efficient trapping and retention of mother cells, determination of the replicative lifespan, and tracking of yeast cells throughout their entire lifespan. Using the high-resolution and large-scale data generated from the high-throughput yeast aging analysis (HYAA) chips, we investigated particular longevity-related changes in cell morphology and characteristics, including critical cell size, terminal morphology, and protein subcellular localization. In addition, because of the significantly improved retention rate of yeast mother cell, the HYAA-Chip was capable of demonstrating replicative lifespan extension by calorie restriction.

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