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Studying the role of buffers in impedance spectroscopy is a relatively unexplored area. We demonstrate a special class of biologically relevant buffers known as Good's zwitterionic buffers that show improved performance over standard electrolyte buffers (e.g. PBS) currently widely used in impedance spectroscopy measurements of bacterial suspensions. Our theoretical and experimental comparisons of conductivity of classical and zwitterionic buffers at various different concentrations show that ion-ion interaction effects are significantly higher in zwitterionic buffers as compared to classical buffers at the concentrations at which they are used. This and the fact that zwitterions have larger sizes leads to the lowering of their conductivity which significantly improves their impedance sensing ability. We illustrate through an example of heat-induced ionic release in model S. typhi and S. aureus bacteria that having a low conductivity buffer is indeed beneficial for biological impedance measurements. In fact, the best buffer for impedance studies can be chosen solely based on their electrical properties as long as they are also biologically compatible. This gives Good's zwitterionic buffers an edge over conventional media as they satisfy both these criteria.

Antimicrobial resistance (AMR) is caused by inappropriate or excessive antibiotic consumption. Early diagnosis of bacterial infections can greatly curb empirical treatment and thus AMR. Current diagnostic procedures are time-consuming as they rely on gene amplification and cell culture techniques that are inherently limited by the doubling rate of the involved species. Further, biochemical methods for species identification and antibiotic susceptibility testing for drug/dose effectiveness take several days and are non-scalable. We report a real-time, label-free approach called DEPIS that combines dielectrophoresis (DEP) for bacterial enrichment and impedance spectroscopy (IS) for cell viability analysis under 60 min. Target bacteria are captured on interdigitated electrodes using DEP (30 min) and their antibiotic-induced stress response is measured using IS (another 30 min). This principle is used to generate minimum bactericidal concentration (MBC) plots by measuring impedance change due to ionic release by dying bacteria in a low conductivity buffer. The results are rapid since they rely on cell death rather than cell growth which is an intrinsically slower process. The results are also highly specific and work across all bactericidal antibiotics studied, irrespective of their cellular target or drug action mechanism. More importantly, preliminary results with clinical isolates show that methicillin-susceptible Staphylococcus aureus (MSSA) can easily be differentiated from methicillin-resistant S. aureus (MRSA) under 1 h. This rapid cell analyses approach can aid in faster diagnosis of bacterial infections and benefit the clinical decision-making process for antibiotic treatment, addressing the critical issue of AMR.