Double-peak signal features in microfluidic impedance flow cytometry enable sensitive measurement of cell membrane...

The probing of individual cells at specific frequency regimes in a microfluidic impedance flow cytometer led to the observation of unusual "double peak" features in the reactive component of the resulting signal. The phenomenon was restricted to the lower frequencies (400-800 kHz) of the β-dispersion regime and its occurrence was facilitated by the co-planar microelectrode geometry in the device. To understand the reasons for this anomalous behaviour, the system was modelled using COMSOL. The simulated model agreed well with experimental observations and provided insight into the origins of this signal profile and the effect of various parameters on its behaviour. One of the most significant observations of this study was the high sensitivity of the features in the "double peak" profile to changes in cell membrane capacitance (CMC), compared to conventional "single peaks" of reactive impedance. This was consequently exploited to accurately distinguish populations of normal and glutaraldehyde treated erythrocytes based on variations in their CMC, indicating a drastic decrease in the CMC of treated cells. Additionally, we demonstrate the applicability of using this double peak effect to identify cell populations within a mixture of PBMCs. This study is an improvement over conventional approaches of measuring CMC via impedance flow cytometry by enabling the measurement of both cell size and cell membrane properties at a single frequency rather than using multiple frequencies. Using a single frequency significantly simplifies the system and reduces the associated costs. Additionally, this technique enables the measurement of CMC at relatively low frequencies.