Dual Fluorochrome Flow Cytometric Assessment of Yeast Viability | Zendy

Bradley J. Hernlem | Zendy; Sui-Sheng T. Hua | Zendy

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Dual Fluorochrome Flow Cytometric Assessment of Yeast Viability

Author(s) -

Bradley J. Hernlem,

Sui-Sheng T. Hua

Publication year - 2010

Publication title -

current microbiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.578

H-Index - 90

eISSN - 1432-0991

pISSN - 0343-8651

DOI - 10.1007/s00284-009-9576-7

Subject(s) - yeast , calcein , biology , esterase , flow cytometry , viability assay , biochemistry , vital stain , staining , cell sorting , stain , cell , microbiology and biotechnology , enzyme , membrane , genetics

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 DiBAC(4)(5) (Bis-(1,3-dibutylbarbituric acid)pentamethine oxonol), a structural analog of the commonly used DiBAC(4)(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.

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