High-Throughput Rapid Yeast Chronological Lifespan Assay

Methods Mol Biol. 2021:2196:229-233. doi: 10.1007/978-1-0716-0868-5_18.

Abstract

The budding yeast is a valuable model system for discovering molecular mechanisms underlying cellular aging. This is due to the ease of performing genetic manipulations in yeast and the vast number of evolutionarily conserved genes that have been found to regulate cellular health and lifespan from yeast to humans. Lifespan assays are an essential tool for examining the effects of these genes on longevity. There are two ways lifespan is measured in yeast: replicative lifespan (RLS) and chronological lifespan (CLS). RLS is a measure of how many divisions an individual mother cell will undergo. CLS measures the length of time nondividing cells survive. Previously described CLS assays involved diluting and plating cells of a culture and counting the colonies that arose. While effective, this method is both time and labor intensive. Here, we describe a method for a high-throughput rapid CLS assay that is both time- and cost-efficient.

Keywords: Brewing yeast; Chronological lifespan (CLS); High throughput; MTT viability assay.

MeSH terms

  • Biological Assay
  • Cellular Senescence*
  • Data Analysis
  • High-Throughput Screening Assays*
  • Longevity*
  • Saccharomyces cerevisiae / physiology
  • Tetrazolium Salts
  • Thiazoles
  • Yeasts / physiology*

Substances

  • Tetrazolium Salts
  • Thiazoles
  • thiazolyl blue