Effect of diffusion on impedance measurements in a hydrodynamic flow focusing sensor

This paper investigated the effects of diffusion between non-conductive sheath and conductive sample fluids in an impedance-based biosensor. Impedance measurements were made with 2- and 4-electrode configurations. The 4-electrode design offers the advantage of impedance measurements at low frequencies (<1 kHz) without the deleterious effects of double layer impedance which are present in the 2-electrode design. Hydrodynamic flow focusing was achieved with a modified T-junction design with a smaller cross-section for the sample channel than for the focusing channel, which resulted in 2D focusing of the sample stream with just one sheath stream. By choosing a non-conductive sheath fluid and a conductive sample fluid, the electric field was confined to the focused stream. In order to utilize this system for biosensing applications, we characterized it for electrical and flow parameters. In particular, we investigated the effects of varying flow velocities and flow-rate ratios on the focused stream. Increasing flow-rate ratios reduced the cross-sectional area of the focused streams as was verified by finite element modeling and confocal microscopy. Antibody mediated binding of Escherichia coli to the electrode surface caused an increase in solution resistance at low frequencies. The results also showed that the diffusion mass transport at the interface of the two streams limited the benefits of increased flow focusing. Increasing flow velocities could be used to offset the diffusion effect. To optimize detection sensitivity, flow parameters and mass transport must be considered in conjunction, with the goal of reducing diffusion of conducting species out of the focused stream while simultaneously minimizing its cross-sectional area.