There is an urgent need to develop simple and fast antimicrobial susceptibility tests (ASTs) that allow informed prescribing of antibiotics. Here, we describe a label-free AST that can deliver results within an hour, using an actively dividing culture as starting material. The bacteria are incubated in the presence of an antibiotic for 30 min, and then approximately 105 cells are analysed one-by-one with microfluidic impedance cytometry for 2–3 min. The measured electrical characteristics reflect the phenotypic response of the bacteria to the mode of action of a particular antibiotic, in a 30-minute incubation window. The results are consistent with those obtained by classical broth microdilution assays for a range of antibiotics and bacterial species.

Single cell impedance cytometry is a label free analysis technique that is now widely used to measure the electrical properties of cell, and to differentiate different subpopulations. Current techniques are limited to measuring the impedance of a single cell at one or two simultaneous frequencies. Also there are no methods that extrapolate the intrinsic electrical properties of single cells. We demonstrate a new approach that uses multi-frequency impedance measurements to determine the complete intrinsic electrical properties of thousands of single cells at high throughput. The applicability of the method is demonstrated by measuring the properties of red blood cells and red cell ghosts, deriving the unique values of conductivity and permittivity of the membrane and cytoplasm for each individual cell.

Single cell impedance cytometry is a label-free electrical analysis method that requires minimal sample preparation and has been used to count and discriminate cells on the basis of their impedance properties. This paper shows experimental and numerically simulated impedance signals for test particles (6 μm diameter polystyrene) flowing through a microfluidic channel. The variation of impedance signal with particle position is mapped using numerical simulation and these results match closely with experimental data. We demonstrate that for a nominal 40 μm × 40 μm channel, the impedance signal is independent of position over the majority of the channel area, but shows large experimentally verifiable variation at extreme positions. The parabolic flow profile in the channel ensures that most of the sample flows through the area of uniform signal. At high flow rates inertial focusing is observed; the particles flow in equal numbers through two equilibrium positions reducing the coefficient of variance (CV) in the impedance signals to negligible values.

This paper describes a new design of microfluidic impedance cytometer enabling accurate characterization of particles without the need for focusing. The approach uses multiple pairs of electrodes to measure the transit time of particles through the device using two simultaneous different current measurements, a transverse (top to bottom) current and an oblique current. This gives a new metric that can be used to estimate the vertical position of the particle trajectory through the microchannel. This parameter effectively compensates for the non-uniform electric field in the channel that is an unavoidable consequence of the use of planar parallel facing electrodes. The new technique is explained and validated using numerical modelling. Impedance data for 5, 6 and 7 µm particles are collected and compared with simulations. The method gives excellent Coefficient of Variation in (electrical) radius of particles of 1% for a sheath less configuration.