Department of Chemical and Biomolecular Engineering, University of Houston, Houston, TX, USA.
Methods Mol Biol. 2021;2357:209-222. doi: 10.1007/978-1-0716-1621-5_14.
Persister cells are defined as a small fraction of phenotypic variants in a cell population that are temporarily tolerant to bactericidal antibiotics. Persisters are not mutant cells; they generally survive lethal concentrations of antibiotics due to their transient nongrowing state. Persister cells have the ability to resuscitate after the end of antibiotic treatment. Despite significant advancements in the understanding of the molecular mechanisms underlying persister formation, we still have little information about their resuscitation mechanisms. In this chapter, we describe a method to detect and monitor persister resuscitation at the single-cell level using flow cytometry analysis. This method enables us to not only assess the resuscitation characteristics of persisters but also determine and quantify various subpopulations in antibiotic-treated cultures, including viable but nonculturable (VBNC) and dead cells.
持留细胞被定义为细胞群体中表型变异的一小部分，它们暂时耐受杀菌抗生素。持留细胞不是突变细胞；它们通常由于其短暂的非生长状态而在致死浓度的抗生素下存活。持留细胞在抗生素治疗结束后有能力复苏。尽管在理解持留形成的分子机制方面取得了重大进展，但我们对它们的复苏机制仍然知之甚少。在本章中，我们描述了一种使用流式细胞术分析在单细胞水平上检测和监测持留细胞复苏的方法。该方法不仅使我们能够评估持留细胞的复苏特征，还能够确定和量化抗生素处理培养物中的各种亚群，包括活但不可培养（VBNC）和死亡细胞。
