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

Myeong Chan Jo, Wei Liu, Liang Gu, Weiwei Dang, Lidong Qin

Research output: Contribution to journalArticle

77 Scopus citations

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 highthroughput 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.

Original languageEnglish (US)
Pages (from-to)9364-9369
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume112
Issue number30
DOIs
StatePublished - Jul 28 2015

Keywords

  • Calorie restriction
  • High-throughput
  • Microfluidics
  • Replicative aging
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • General

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