Porter J, Pickup R, Edwards C
Department of Genetics and Microbiology, University of Liverpool, UK.
FEMS Microbiol Lett. 1995 Oct 15;132(3):259-62. doi: 10.1016/0378-1097(95)00320-5.
The ionophore, valinomycin, was investigated as a possible means of bacterial viability assessment using the fluorescent membrane potential dye rhodamine 123. Membrane hyperpolarisation in Escherichia coli, Pseudomonas fluorescens, Enterobacter aerogenes and Arthrobacter globiformis was examined during exponential growth and during stress by brief starvation in a high sodium, low potassium buffer using flow cytometric analysis of rhodamine 123 uptake. Dye uptake was variable both between species and amongst cells from the same culture. Exponential phase cells showed no increase in dye uptake due to valinomycin treatment. Stressed P. fluorescens cells responded to valinomycin treatment by increased dye uptake, while stressed E. coli and A. globiformis cells showed no response. Approximately 50% of stressed Eb. aerogenes cells responded to valinomycin. The results demonstrate the limitations of rhodamine dye for viability analysing the viability of diverse bacterial communities and underline the degree of cell heterogeneity in batch cultures.
研究了离子载体缬氨霉素作为一种使用荧光膜电位染料罗丹明123评估细菌活力的可能方法。通过对罗丹明123摄取进行流式细胞术分析，在指数生长期以及在高钠低钾缓冲液中短暂饥饿应激期间，检测了大肠杆菌、荧光假单胞菌、产气肠杆菌和球形节杆菌中的膜超极化。染料摄取在不同物种之间以及来自同一培养物的细胞之间存在差异。指数生长期的细胞经缬氨霉素处理后染料摄取没有增加。应激的荧光假单胞菌细胞对缬氨霉素处理的反应是染料摄取增加，而应激的大肠杆菌和球形节杆菌细胞没有反应。约50%应激的产气肠杆菌细胞对缬氨霉素有反应。结果证明了罗丹明染料在分析不同细菌群落活力方面的局限性，并强调了分批培养中细胞异质性的程度。
