A high-throughput method for quantifying metabolically active yeast cells

Subir Kumar Nandy, Peter Boldsen Knudsen, Alexander Rosenkjær, Anna Eliasson Lantz, Jette Thykær, Mhairi Workman

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Abstract

By redesigning the established methylene blue reduction test for bacteria and yeast, we present a cheap and efficient methodology for quantitative physiology of eukaryotic cells applicable for high-throughput systems. Validation of themethod in fermenters and highthroughput systems proved equivalent, displaying reduction curves that interrelated directly with CFU counts. For growth rate estimation, the methylene blue reduction test (MBRT) proved superior, since the discriminatory nature of the method allowed for the quantification of metabolically active cells only, excluding dead cells. The drop in metabolic activity associated with the diauxic shift in yeast proved more pronounced for the MBRT-derived curve compared with OD curves, consistent with a dramatic shift in the ratio between live and dead cells at this metabolic event. This method provides a tool with numerous applications, e.g. characterizing the death phase of stationary phase cultures, or in drug screens with pathogenic yeasts.
Original languageEnglish
JournalYeast
Volume32
Issue number6
Pages (from-to)461-468
Number of pages8
ISSN0749-503X
DOIs
Publication statusPublished - 2015

Keywords

  • Saccharomyces cerevisiae
  • Metabolically active cells
  • Quantification
  • High throughput
  • Methylene blue

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