An investigation of hydroethidine as a fluorescent vital stain for prokaryotes

Hydroethidine (HE), manufactured by the chemical reduction of ethidium bromide (EB), was found to be decomposed to a bright red-fluorescent product by a range of microorganisms. Therefore, HE seemed to have potential as a fluorescent vital stain for microorganisms. However, inhibited or killed bacterial cultures still fluoresced red. Short periods (2–4 min) of excitation of sterile solutions of HE with a UV light yielded an orange/red product. Thus, HE seems to be decomposed biologically by a number of microorganisms but also abiotically by UV light-mediated process to red fluorescent material. These observations suggest that the oxidation of HE has only limited potential for assessing microbial activity.