The single-entity impact method is a very sensitive and powerful electroanalytical tool for biological and environmental purposes such as electrochemical detection of pathogens.[1] The versatility of ultramicroelectrodes (UMEs) gives unique and essential properties for electroanalytical sensing owing to their small size, high sensitivity, fast steady-state response, low double-layer charging current and minimal ohmic loss. The observation of these stochastic events can provide information on various single nanoparticles contrary to ensemble (bulk) measurements. The main advantage of studying collisions of single entities is the low limit of detection (in principle, one single species) inherent to this electroanalytical method and the ability to study single entities (cells, viruses, nanoparticles...) in real time (dynamic measurement).[2–4]Electrochemistry of single redox liposome impacts at an UME consists in detecting the electrolysis of a redox probe encapsulated inside a liposome when it is released at the UME after impact (or collision). The UME is polarized at the oxidation or reduction potential of the encapsulated redox probe and a concentration of several pico-molar of redox liposomes added to the aqueous buffer electrolyte is enough to detect “current spikes” in the chronoamperometry (i-t) curve corresponding to discrete collision events.[3,4] The electron transfer does not readily occur across a lipid bilayer, thus the electrolysis of the liposome redox active content after collision and membrane rupture or opening at the UME surface led to many studies dealing with the membrane permeation mechanism.[3,4]Here, our work is based on the previous results where no current spike was observed in the chronoamperometry curve because the redox DMPC liposomes did not break during impact (or collision) onto the UME surface.[3,4] Hence, the electrochemical sensing principle is based on the weakening of the liposomes lipid membrane upon interaction with a destructive bacterial virulence factor which leads upon impact at an UME to the breakdown of the liposomes and the release/electrolysis of its encapsulated redox probe, as previously reported.[5] In the presence of RL toxin in solution (acting like a surfactant in the lipid membrane), current spikes corresponding to the electrolysis of the encapsulated redox probe released from weakened liposomes are detected (see Figure). Thanks to the redox liposome single impact electrochemistry, the highest detection limit of RL toxin (500 nM) has been reached in comparison to several micromoles per liter previously reported.[5][1] Lebègue, E.; Costa, N.L.; Louro, R.O.; Barrière, F. Communication—Electrochemical Single Nano-Impacts of Electroactive Shewanella Oneidensis Bacteria onto Carbon Ultramicroelectrode. J. Electrochem. Soc. 2020, 167, 105501, doi:10.1149/1945-7111/ab9e39.[2] Dick, J.E.; Lebègue, E.; Strawsine, L.M.; Bard, A.J. Millisecond Coulometry via Zeptoliter Droplet Collisions on an Ultramicroelectrode. Electroanalysis 2016, 28, 2320–2326, doi:10.1002/elan.201600182.[3] Lebègue, E.; Anderson, C.M.; Dick, J.E.; Webb, L.J.; Bard, A.J. Electrochemical Detection of Single Phospholipid Vesicle Collisions at a Pt Ultramicroelectrode. Langmuir 2015, 31, 11734–11739, doi:10.1021/acs.langmuir.5b03123.[4] Lebègue, E.; Barrière, F.; Bard, A.J. Lipid Membrane Permeability of Synthetic Redox DMPC Liposomes Investigated by Single Electrochemical Collisions. Anal. Chem. 2020, 92, 2401–2408, doi:10.1021/acs.analchem.9b02809.[5] Luy, J.; Ameline, D.; Thobie-Gautier, C.; Boujtita, M.; Lebègue, E. Detection of Bacterial Rhamnolipid Toxin by Redox Liposome Single Impact Electrochemistry. Angew. Chem. Int. Ed. 2021, accepted , doi: 10.1002/anie.202111416. Figure 1