Current Microbiology

, Volume 61, Issue 1, pp 57–63

Dual Fluorochrome Flow Cytometric Assessment of Yeast Viability

Authors

    • Agricultural Research Service, Western Regional Research Center, Foodborne Contaminants Research UnitU.S. Department of Agriculture
  • Sui-Sheng Hua
    • Agricultural Research Service, Western Regional Research Center, Plant Mycotoxins Research UnitU.S. Department of Agriculture
Article

DOI: 10.1007/s00284-009-9576-7

Cite this article as:
Hernlem, B. & Hua, S. Curr Microbiol (2010) 61: 57. doi:10.1007/s00284-009-9576-7

Abstract

A novel staining protocol is reported for the assessment of viability in yeast, specifically the biocontrol yeast, Pichia anomala. Employing both the red fluorescent membrane potential sensitive oxonol stain DiBAC4(5) (Bis-(1,3-dibutylbarbituric acid)pentamethine oxonol), a structural analog of the commonly used DiBAC4(3) (Bis-(1,3-dibutylbarbituric acid)trimethine oxonol), with one of the esterase dependent green fluorogenic probes such as CFDA-AM (5-Carboxyfluorescein diacetate, acetoxymethyl ester) or Calcein-AM (Calcein acetoxymethyl ester), a two-color flow cytometric method was developed, which yields rapid quantitative information on the vitality and vigor of yeast cell cultures. The method was validated by cell sorting and analysis of live, heat killed, and UV-treated yeast.

Copyright information

© US Government 2010