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Electrokinetic (EK) microsystems, which are capable of performing separations without the need for labeling analytes, are a rapidly growing area in microfluidics. The present work demonstrated three distinct binary microbial separations, computationally modeled and experimentally performed, in an insulator-based EK (iEK) system stimulated by DC-biased AC potentials. The separations had an increasing order of difficulty. First, a separation between cells of two distinct domains (Escherichia coli and Saccharomyces cerevisiae) was demonstrated. The second separation was for cells from the same domain but different species (Bacillus subtilis and Bacillus cereus). The last separation included cells from two closely related microbial strains of the same domain and the same species (two distinct S. cerevisiae strains). For each separation, a novel computational model, employing a continuous spatial and temporal function for predicting the particle velocity, was used to predict the retention time (tR,p) of each cell type, which aided the experimentation. All three cases resulted in separation resolution values Rs>1.5, indicating complete separation between the two cell species, with good reproducibility between the experimental repetitions (deviations < 6%) and good agreement (deviations < 18%) between the predicted tR,p and experimental (tR,e) retention time values. This study demonstrated the potential of DC-biased AC iEK systems for performing challenging microbial separations. 

This study proposes a strategy for improving the separation resolution of binary microparticle mixtures by modifying the post array arrangement in insulator-based electrokinetic devices. 

There is a rising need for rapid and reliable analytical methods for separating microorganisms in clinical and biomedical applications. Microscale-insulator-based electrokinetic (iEK) systems have proven to be robust platforms for assessing a wide variety of microorganisms. Traditionally, iEK systems are usually stimulated with direct-current (DC) potentials. This work presents a comparison between using DC potentials and using DC-biased alternating-current (AC) potentials in iEK systems for the separation of microorganisms. The present study, which includes mathematical modeling and experimentation, compares the separation of bacterial and yeast cells in two distinct modes by using DC and DC-biased AC potentials. The quality of both separations, assessed in terms of separation resolution (Rs), showed a complete separation (Rs = 1.51) with the application of a DC-biased low-frequency AC signal but an incomplete separation (Rs = 0.55) with the application of an RMS-equivalent DC signal. Good reproducibility between experimental repetitions (<10%) was obtained, and good agreement (~18% deviation) was observed between modeling and experimental retention times. The present study demonstrates the potential of extending the limits of iEK systems by employing DC-biased AC potentials to perform discriminatory separations of microorganisms that are difficult to separate with the application of DC potentials. 

Insulator-based microfluidic devices are attractive for handling biological samples due to their simple fabrication, low-cost, and efficiency in particle manipulation. However, their widespread application is limited by the high operation voltages required to achieve particle trapping. We present a theoretical, numerical, and experimental study that demonstrates these voltages can be significantly reduced (to sub-100 V) in direct-current insulator-based electrokinetic (DC-iEK) devices for micron-sized particles. To achieve this, we introduce the concept of the amplification factor—the fold-increase in electric field magnitude due to the presence of an insulator constriction—and use it to compare the performance of different microchannel designs and to direct our design optimization process. To illustrate the effect of using constrictions with smooth and sharp features on the amplification factor, geometries with circular posts and semi-triangular posts were used. These were theoretically approximated in two different systems of coordinates (bipolar and elliptic), allowing us to provide, for the first time, explicit electric field amplification scaling laws. Finite element simulations were performed to approximate the 3D insulator geometries and provide a parametric study of the effect of changing different geometrical features. These simulations were used to predict particle trapping voltages for four different single-layer microfluidic devices using two particle suspensions (2 and 6.8 μm in size). The general agreement between our models demonstrates the feasibility of using the amplification factor, in combination with nonlinear electrokinetic theory, to meet the prerequisites for the development of portable DC-iEK microfluidic systems.