Dual fluorochrome flow cytometric assessment of yeast viability.
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.
Hernlem
Bradley
B
Agricultural Research Service, Western Regional Research Center, Foodborne Contaminants Research Unit, U.S. Department of Agriculture, 800 Buchanan St., Albany, CA 94710, USA. bradley.hernlem@ars.usda.gov
Hua
Sui-Sheng
SS
D001463:Barbiturates; D015169:Colony Count, Microbial; D005434:Flow Cytometry / Q000379:methods*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007555:Isoxazoles; D008564:Membrane Potentials; D050296:Microbial Viability; D010843:Pichia / Q000254:growth & development / Q000502:physiology* / Q000528:radiation effects; D013194:Staining and Labeling; D014466:Ultraviolet Rays
C505935:5-carboxyfluorescein diacetate acetoxymethyl ester; D001463:Barbiturates; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007555:Isoxazoles; C096530:bis(1,3-dibutylbarbituric acid)pentamethine oxonol; C085925:calcein AM