Agricultural Research Service, Western Regional Research Center, Foodborne Contaminants Research Unit, U.S. Department of Agriculture, 800 Buchanan St., Albany, CA 94710, USA.
Curr Microbiol. 2010 Jul;61(1):57-63. doi: 10.1007/s00284-009-9576-7. Epub 2010 Jan 5.
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.
报道了一种用于评估酵母（特别是生防酵母异常毕赤酵母）活力的新型染色方案。该方案同时使用红色荧光膜电位敏感型吖啶橙染料 DiBAC(4)(5)（双（1,3-二丁基巴比妥酸）五甲川菁氧鎓）和酯酶依赖性绿色荧光探针，如 CFDA-AM（5-羧基荧光素二乙酸酯，乙酰氧基甲酯）或 Calcein-AM（钙黄绿素乙酰氧基甲酯），开发了一种双色流式细胞术方法，可快速定量获得酵母细胞培养物的活力和活力信息。该方法通过细胞分选和对活细胞、热致死细胞和 UV 处理细胞的分析进行了验证。
