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Antibiotics such as the quinolones and fluoroquinolones kill bacterialpathogens ultimately through DNA damage. They target the essential type IIAtopoisomerases in bacteria by stabilising the normally transient double strandbreak state which is created to modify the supercoiling state of the DNA. Herewe discuss the development of these antibiotics and their method of action.Existing methods for DNA damage visualisation, such as the comet assay andimmunofluorescence imaging can often only be analysed qualitatively and thisanalysis is subjective. We describe a putative single-molecule fluorescencetechnique for quantifying DNA damage via the total fluorescence intensity of aDNA origami tile fully saturated with an intercalating dye, along with theoptical requirements for how to implement these into a light microscopy imagingsystem capable of single-molecule millisecond timescale imaging. This systempromises significant improvements in reproducibility of the quantification ofDNA damage over traditional techniques.

Designing a single-molecule biophysics tool for characterizing DNA damage for techniques that kill infectious pathogens through DNA damage effects