Observing Bacterial Persistence at Single-Cell Resolution

Methods Mol Biol. 2021;2357:85-93. doi: 10.1007/978-1-0716-1621-5_6.


Within a bacterial population, there can be a subpopulation of cells with an antibiotic-tolerant persister phenotype characterized by long lag phase. Their long lag phase necessitates long (hours or days) periods of single-cell observation to capture high-quality quantitative information about persistence. We describe a method of single-cell imaging using glass bottom dishes and a nutrient agarose pad that allows for long-term single-cell microscopy observation in a stable environment. We apply this method to characterize the lag phase and persistence of individual Escherichia coli cells.

Keywords: Antibiotic resistance; Lag time; Persistence; Persisters; Single-cell microscopy.

MeSH terms

  • Anti-Bacterial Agents
  • Bacteria*
  • Escherichia coli
  • Escherichia coli Infections
  • Escherichia coli Proteins
  • Humans


  • Anti-Bacterial Agents
  • Escherichia coli Proteins