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1. Determination of the Empirical Electrokinetic Equilibrium Condition of Microorganisms in Microfluidic Devices

   https://doi.org/10.3390/bios10100148  

   Coll De Peña, Adriana; Hill, Nicole; Lapizco-Encinas, Blanca H. (October 2020, Biosensors)

   null (Ed.)

   The increased concern regarding emerging pathogens and antibiotic resistance has drawn interest in the development of rapid and robust microfluidic techniques to analyze microorganisms. The novel parameter known as the electrokinetic equilibrium condition (EEEC) was presented in recent studies, providing an approach to analyze microparticles in microchannels employing unique electrokinetic (EK) signatures. While the EEEC shows great promise, current estimation approaches can be time-consuming or heavily user-dependent for accurate values. The present contribution aims to analyze existing approaches for estimating this parameter and modify the process into an accurate yet simple technique for estimating the EK behavior of microorganisms in insulator-based microfluidic devices. The technique presented here yields the parameter called the empirical electrokinetic equilibrium condition (eEEEC) which works well as a value for initial approximations of trapping conditions in insulator-based EK (iEK) microfluidic systems. A total of six types of microorganisms were analyzed in this study (three bacteria and three bacteriophages). The proposed approach estimated eEEEC values employing images of trapped microorganisms, yielding high reproducibility (SD 5.0–8.8%). Furthermore, stable trapping voltages (sTVs) were estimated from eEEEC values for distinct channel designs to test that this parameter is system-independent and good agreement was obtained when comparing estimated sTVs vs. experimental values (SD 0.3–19.6%). The encouraging results from this work were used to generate an EK library of data, available on our laboratory website. The data in this library can be used to design tailored iEK microfluidic devices for the analysis of microorganisms. 

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2. Fine-Tuning Electrokinetic Injections Considering Nonlinear Electrokinetic Effects in Insulator-Based Devices

   https://doi.org/10.3390/mi12060628  

   Miller, Abbi; Hill, Nicole; Hakim, Kel; Lapizco-Encinas, Blanca H. (June 2021, Micromachines)

   null (Ed.)

   The manner of sample injection is critical in microscale electrokinetic (EK) separations, as the resolution of a separation greatly depends on sample quality and how the sample is introduced into the system. There is a significant wealth of knowledge on the development of EK injection methodologies that range from simple and straightforward approaches to sophisticated schemes. The present study focused on the development of optimized EK sample injection schemes for direct current insulator-based EK (DC-iEK) systems. These are microchannels that contain arrays of insulating structures; the presence of these structures creates a nonuniform electric field distribution when a potential is applied, resulting in enhanced nonlinear EK effects. Recently, it was reported that the nonlinear EK effect of electrophoresis of the second kind plays a major role in particle migration in DC-iEK systems. This study presents a methodology for designing EK sample injection schemes that consider the nonlinear EK effects exerted on the particles being injected. Mathematical modeling with COMSOL Multiphysics was employed to identify proper voltages to be used during the EK injection process. Then, a T-microchannel with insulating posts was employed to experimentally perform EK injection and separate a sample containing two types of similar polystyrene particles. The quality of the EK injections was assessed by comparing the resolution (Rs) and number of plates (N) of the experimental particle separations. The findings of this study establish the importance of considering nonlinear EK effects when planning for successful EK injection schemes. 

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3. Assessing the Discriminatory Capabilities of iEK Devices under DC and DC-Biased AC Stimulation Potentials

   https://doi.org/10.3390/mi14122239  

   Nasir_Ahamed, Nuzhet Nihaar; Mendiola-Escobedo, Carlos A; Perez-Gonzalez, Victor H; Lapizco-Encinas, Blanca H (December 2023, Micromachines)

   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. 

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4. Dielectrophoretic trapping of particles flowing through an array of conductive cylinders

   https://doi.org/10.1063/5.0305604  

   Mondal, Tonoy K; Williams, Stuart J (December 2025, Physics of Fluids)

   Dielectrophoresis (DEP) is a label-free electrokinetic method for selectively trapping polarizable particles using non-uniform electric fields. While co-planar electrode systems are common, their inherent DEP force distribution limits throughput. This study presents a computationally efficient framework for modeling two-dimensional DEP-based particle trapping in ordered arrays of conductive cylinders. These cylinders are modeled at a range of sizes, from micrometers to nanometers, to represent microfluidic systems consisting of conductive pillars, nanofibers, etc. Analytical solutions for fluid flow and electric potential were derived using eigenfunction expansions and collocation, then used in a particle tracking model that includes hydrodynamic drag, Brownian motion, and multipolar DEP forces. Although focused on conductive arrays, this framework is extensible to other configurations. This work provides a foundation for future work in the design of high-throughput DEP systems. Both dimensionless and dimensional analyses were performed across a wide range of particle sizes (30 nm to 3 μm), voltages (10 mV to 100 V), and array geometries. No specific optimal cylinder size was found; instead, optimal performance arises from a balance between DEP force distribution and flow through the cylinder array gap. Diamond-oriented arrays exhibited enhanced trapping under moderate dielectrophoretic velocity-to-fluid velocity ratios (up to 39% greater), while square arrays performed better under low-field and large-cylinder conditions (up to 40% greater). 

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5. Switching Separation Migration Order by Switching Electrokinetic Regime in Electrokinetic Microsystems

   https://doi.org/10.3390/BIOS14030119  

   Vaghef-Koodehi, Alaleh; Lapizco-Encinas, Blanca H (March 2024, Biosensors)

   Analyte migration order is a major aspect in all migration-based analytical separations methods. Presented here is the manipulation of the migration order of microparticles in an insulator-based electrokinetic separation. Three distinct particle mixtures were studied: a binary mixture of particles with similar electrical charge and different sizes, and two tertiary mixtures of particles of distinct sizes. Each one of the particle mixtures was separated twice, the first separation was performed under low voltage (linear electrokinetic regime) and the second separation was performed under high voltage (nonlinear electrokinetic regime). Linear electrophoresis, which discriminates particles by charge, is the dominant electrokinetic effect in the linear regime; while nonlinear electrophoresis, which discriminates particles by size and shape, is the dominant electrokinetic effect in the nonlinear regime. The separation results obtained with the three particle mixtures illustrated that particle elution order can be changed by switching from the linear electrokinetic regime to the nonlinear electrokinetic regime. Also, in all cases, better separation performances in terms of separation resolution (Rs) were obtained by employing the nonlinear electrokinetic regime allowing nonlinear electrophoresis to be the discriminatory electrokinetic mechanism. These findings could be applied to analyze complex samples containing bioparticles of interest within the micron size range. This is the first report where particle elution order is altered in an iEK system. 

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