We present a method capable of detecting single slow-growing and growth-arrested cells in a bacterial culture composed of physiologically and phenotypically different cells. Unlike the use of transcriptional reporters to gauge the metabolic activities in cells, here, we fuse two different fluorescent proteins with distinctive maturation rates to construct a timer to directly determine the growth rate of single Pseudomonas aeruginosa cells. We demonstrate that the dual-color fluorescent timer can indicate the slow-growing and growth-arrested cells from bacterial cultures in the presence of various environmental stresses, including nutrient starvation or antibiotic treatments, which greatly expand the methods for detecting and isolating persister cells.

中文翻译：

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双色荧光计时器可检测被阻止的致病细菌

我们提出了一种能够在由生理和表型不同的细胞组成的细菌培养物中检测单个缓慢生长和生长停滞的细胞的方法。与使用转录报告子评估细胞中的代谢活性不同，在这里，我们将具有不同成熟率的两种不同荧光蛋白融合在一起，以构建一个计时器来直接确定单个铜绿假单胞菌细胞的生长速率。我们证明了双色荧光计时器可以指示在各种环境压力（包括营养缺乏或抗生素治疗）的存在下细菌培养的缓慢生长和生长受阻的细胞，这极大地扩展了检测和分离持久性细胞的方法。