Label-Free Detection of Single Living Bacteria: Single-Entity Electrochemistry Targeting Metabolic Products

This study presents a novel approach employing single-entity electrochemistry for the label-free detection of living Escherichia coli. By examination of the collision signals generated from the reduction of hydrogen peroxide, a metabolic product of E. coli that accumulates on the cell surface, the concentration of living bacteria can be determined. Within a broad concentration range from 3.0 × 10⁷ to 1.0 × 10⁹ cells/mL, cell aggregation was not observed. Cell migration in the solution was primarily governed by diffusion, exhibiting a diffusion coefficient of 6.8 × 10^-9 cm²/s. The collision frequency exhibits a linear relationship with the cell concentration, aligning well with theoretical predictions. Through statistical analysis of each collision signal's integrated charge quantity, the metabolic activity of single cells can be assessed. This method was applied to a cytotoxicity assay, where it monitored the decline in living cell numbers and metabolic activities in addition to identifying potential cell damage during antibiotic treatment.