title: High-throughput analysis of yeast replicative aging using a microfluidic system.
abstract: 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.
authors.affiliation: Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030; Department of Cell and Developmental Biology, Weill Cornell Medical College, New York, NY 10065;
authors.forename: Myeong Chan
authors.initials: MC
authors.lastname: Jo
authors.affiliation: Huffington Center on Aging, Baylor College of Medicine, Houston, TX 77030.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030;
authors.forename: Liang
authors.initials: L
authors.lastname: Gu
authors.affiliation: Huffington Center on Aging, Baylor College of Medicine, Houston, TX 77030 lqin@houstonmethodist.org weiwei.dang@bcm.edu.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Dang
authors.affiliation: Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030; Department of Cell and Developmental Biology, Weill Cornell Medical College, New York, NY 10065; lqin@houstonmethodist.org weiwei.dang@bcm.edu.
authors.forename: Lidong
authors.initials: L
authors.lastname: Qin
mesh_terms: D031204:Caloric Restriction; D002453:Cell Cycle; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D020342:Finite Element Analysis; D017353:Gene Deletion; D049452:Green Fluorescent Proteins / Q000737:chemistry; D057166:High-Throughput Screening Assays; D007091:Image Processing, Computer-Assisted; D046228:Microarray Analysis; D044085:Microfluidics; D008853:Microscopy; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000166:cytology*; D059010:Single-Cell Analysis
chemical_list: D004129:Dimethylpolysiloxanes; D049452:Green Fluorescent Proteins; C013830:baysilon
keywords: Saccharomyces cerevisiae; calorie restriction; high-throughput; microfluidics; replicative aging