Manipulating the insulating post arrangement in DC-biased AC-iEK devices to improve microparticle separations.

There is a growing interest in the advancement of microscale electrokinetic (EK) systems for biomedical and clinical applications, as these systems offer attractive characteristics such as portability, robustness, low sample requirements and short response time. The present work is focused on manipulating the characteristics of the insulating post arrangement in insulator-based EK (iEK) systems for separating a binary mixture of spherical microparticles with same diameter (5.1 µm), same shape, made from the same substrate material and only differing in their zeta potential by ∼14 mV. This study presents a combination of mathematical modeling and experimental separations performed by applying a low-frequency alternating current (AC) voltage in iEK systems with 12 distinct post arrangements. These iEK devices were used to systematically study the effect of three spatial characteristics of the insulating post array on particle separations: the horizontal separation and the vertical separation between posts, and introducing an offset to the posts arrangement. Through normalization of the spatial separation between the insulating posts with respect to particle diameter, guidelines to improve separation resolution for different particle mixtures possessing similar characteristics were successfully identified. The results indicated that by carefully designing the spatial arrangement of the post array, separation resolution values in the range of 1.4-2.8 can be obtained, illustrating the importance and effect of the arrangement of insulating posts on improving particle separations. This study demonstrates that iEK devices, with effectively designed spatial arrangement of the insulating post arrays, have the capabilities to perform discriminatory separations of microparticles of similar characteristics.

Self-Diagnosis of SARS-CoV-2 from Saliva Samples at Home: Isothermal Amplification Enabled by Do-It-Yourself Portable Incubators and Laminated Poly-ethyl Sulfonate Membranes.

COVID-19 made explicit the need for rethinking the way in which we conduct testing for epidemic emergencies. During the COVID-19 pandemic, the dependence on centralized lab facilities and resource-intensive methodologies (e.g., RT-qPCR methods) greatly limited the deployment of widespread testing efforts in many developed and underdeveloped countries. Here, we illustrate the development of a simple and portable diagnostic kit that enables self-diagnosis of COVID-19 at home from saliva samples. We describe the development of a do-it-yourself (DIY) incubator for Eppendorf tubes that can be used to conduct SARS-CoV-2 detection with competitive sensitivity and selectivity from saliva at home. In a proof-of-concept experiment, we assembled Eppendorf-tube incubators at our home shop, prepared a single-tube mix of reagents and LAMP primers in our lab, and deployed these COVID-19 detection kits using urban delivery systems (i.e., Rappifavor or Uber) to more than 15 different locations in Monterrey, México. This straightforward strategy enabled rapid and cost-effective at-home molecular diagnostics of SARS-CoV-2 from real saliva samples with a high sensitivity (100%) and high selectivity (87%).

Trends and challenges in microfluidic methods for protein manipulation-A review.

Proteins are important molecules involved in an immensely large number of biological processes. Being capable of manipulating proteins is critical for developing reliable and affordable techniques to analyze and/or detect them. Such techniques would enable the production of therapeutic agents for the treatment of diseases or other biotechnological applications (e.g., bioreactors or biocatalysis). Microfluidic technology represents a potential solution to protein manipulation challenges because of the diverse phenomena that can be exploited to achieve micro- and nanoparticle manipulation. In this review, we discuss recent contributions made in the field of protein manipulation in microfluidic systems using different physicochemical principles and techniques, some of which are miniaturized versions of already established macro-scale techniques.

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

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.

Fine-Tuning the Characteristic of the Applied Potential To Improve AC-iEK Separations of Microparticles.

There is an immediate need for the development of rapid and reliable methods for microparticle and cell assessments, and electrokinetic (EK) phenomena can be exploited to meet that need in a low cost and label-free fashion. The present study combines modeling and experimentation to separate a binary mixture of microparticles of the same size (5.1 µm), shape (spherical), and substrate material (polystyrene), but with a difference in particle zeta potentials of only ∼14 mV, by applying direct current (DC)-biased low-frequency alternating current (AC) voltages in an insulator-based-EK (iEK) system. Four distinct separations were carried out to systematically study the effect of fine-tuning each of the three main characteristics of the applied voltage: frequency, amplitude, and DC bias. The results indicate that fine-tuning each parameter improved the separation from an initial separation resolution Rs = 0.5 to a final resolution Rs = 3.1 of the fully fine-tuned separation. The separation method exhibited fair reproducibility in retention time with variations ranging from 6 to 26% between experimental repetitions. The present study demonstrates the potential to extend the limits of iEK systems coupled with carefully fine-tuned DC-biased low-frequency AC voltages to perform discriminatory micron-sized particle separations.

Recent advances and challenges in temperature monitoring and control in microfluidic devices.

Temperature is a critical-yet sometimes overlooked-parameter in microfluidics. Microfluidic devices can experience heating inside their channels during operation due to underlying physicochemical phenomena occurring therein. Such heating, whether required or not, must be monitored to ensure adequate device operation. Therefore, different techniques have been developed to measure and control temperature in microfluidic devices. In this contribution, the operating principles and applications of these techniques are reviewed. Temperature-monitoring instruments revised herein include thermocouples, thermistors, and custom-built temperature sensors. Of these, thermocouples exhibit the widest operating range; thermistors feature the highest accuracy; and custom-built temperature sensors demonstrate the best transduction. On the other hand, temperature control methods can be classified as external- or integrated-methods. Within the external methods, microheaters are shown to be the most adequate when working with biological samples, whereas Peltier elements are most useful in applications that require the development of temperature gradients. In contrast, integrated methods are based on chemical and physical properties, structural arrangements, which are characterized by their low fabrication cost and a wide range of applications. The potential integration of these platforms with the Internet of Things technology is discussed as a potential new trend in the field.

Nanomaterials for electrochemical detection of pollutants in water: A review.

The survival of living beings, including humanity, depends on a continuous supply of clean water. However, due to the development of industry, agriculture, and population growth, an increasing number of wastewaters is discarded, and the negative effects of such actions are clear. The first step in solving this situation is the collection and monitoring of pollutants in water bodies to subsequently facilitate their treatment. Nonetheless, traditional sensing techniques are typically laboratory-based, leading to potential diminishment in analysis quality. In this paper, the most recent developments in micro- and nano-electrochemical devices for pollutant detection in wastewater are reviewed. The devices reviewed are based on a variety of electrodes and the sensing of three different categories of pollutants: nutrients and phenolic compounds, heavy metals, and organic matter. From these electrodes, Cu, Co, and Bi showed promise as versatile materials to detect a grand variety of contaminants. Also, the most commonly used material is glassy carbon, present in the detection of all reviewed analytes.

Electrically driven microfluidic platforms for exosome manipulation and characterization.

Exosomes are small extracellular vesicles that can be obtained from several body fluids such as blood and urine. Since these vesicles can carry biomarkers and other cargo, they have application in healthcare diagnostics and therapeutics, such as liquid biopsies and drug delivery. Yet, their identification and separation from a sample remain challenging due to their high degree of heterogeneity and their co-existence with other bioparticles. In this contribution, we review the state-of-the-art on electrical techniques and methods to displace, selectively trap/isolate, and detect/characterize exosomes in microfluidic devices. Although there are many reviews focused on exosome separation using benchtop equipment, such as ultracentrifugation, there are limited reviews focusing on the use of electrical phenomena in microfluidic devices for exosome manipulation and detection. Here, we highlight contributions published during the past decade and present perspectives for this research field for the near future, outlining challenges to address in years to come.

Amplification factor in DC insulator-based electrokinetic devices: a theoretical, numerical, and experimental approach to operation voltage reduction for particle trapping.

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.

Rapid Lipid Content Screening in Neochloris oleoabundans Utilizing Carbon-Based Dielectrophoresis.

In this study, we carried out a heterogeneous cytoplasmic lipid content screening of Neochloris oleoabundans microalgae by dielectrophoresis (DEP), using castellated glassy carbon microelectrodes in a PDMS microchannel. For this purpose, microalgae were cultured in nitrogen-replete (N+) and nitrogen-deplete (N-) suspensions to promote low and high cytoplasmic lipid production in cells, respectively. Experiments were carried out over a wide frequency window (100 kHz-30 MHz) at a fixed amplitude of 7 VPP. The results showed a statistically significant difference between the dielectrophoretic behavior of N+ and N- cells at low frequencies (100-800 kHz), whereas a weak response was observed for mid- and high frequencies (1-30 MHz). Additionally, a finite element analysis using a 3D model was conducted to determine the dielectrophoretic trapping zones across the electrode gaps. These results suggest that low-cost glassy carbon is a reliable material for microalgae classification-between low and high cytoplasmic lipid content-through DEP, providing a fast and straightforward mechanism.

Particle trapping in electrically driven insulator-based microfluidics: Dielectrophoresis and induced-charge electrokinetics.

Electrokinetically driven insulator-based microfluidic devices represent an attractive option to manipulate particle suspensions. These devices can filtrate, concentrate, separate, or characterize micro and nanoparticles of interest. Two decades ago, inspired by electrode-based dielectrophoresis, the concept of insulator-based dielectrophoresis (iDEP) was born. In these microfluidic devices, insulating structures (i.e., posts, membranes, obstacles, or constrictions) built within the channel are used to deform the spatial distribution of an externally generated electric field. As a result, particles suspended in solution experience dielectrophoresis (DEP). Since then, it has been assumed that DEP is responsible for particle trapping in these devices, regardless of the type of voltage being applied to generate the electric field-direct current (DC) or alternating current. Recent findings challenge this assumption by demonstrating particle trapping and even particle flow reversal in devices that prevent DEP from occurring (i.e., unobstructed long straight channels stimulated with a DC voltage and featuring a uniform electric field). The theory introduced to explain those unexpected observations was then applied to conventional "DC-iDEP" devices, demonstrating better prediction accuracy than that achieved with the conventional DEP-centered theory. This contribution summarizes contributions made during the last two decades, comparing both theories to explain particle trapping and highlighting challenges to address in the near future.

A survey of electrokinetically-driven microfluidics for cancer cells manipulation.

Cancer is one of the leading causes of annual deaths worldwide, accounting for nearly 10 million deaths each year. Metastasis, the process by which cancer spreads across the patient's body, is the main cause of death in cancer patients. Because the rising trend observed in statistics of new cancer cases and cancer-related deaths does not allow for an optimistic viewpoint on the future-in relation to this terrible disease-the scientific community has sought methods to enable early detection of cancer and prevent the apparition of metastatic tumors. One such method is known as liquid biopsy, wherein a sample is taken from a bodily fluid and analyzed for the presence of CTCs or other cancer biomarkers (e.g., growth factors). With this objective, interest is growing by year in electrokinetically-driven microfluidics applied for the concentration, capture, filtration, transportation, and characterization of CTCs. Electrokinetic techniques-electrophoresis, dielectrophoresis, electrorotation, and electrothermal and EOF-have great potential for miniaturization and integration with electronic instrumentation for the development of point-of-care devices, which can become a tool for early cancer diagnostics and for the design of personalized therapeutics. In this contribution, we review the state of the art of electrokinetically-driven microfluidics for cancer cells manipulation.

Toward low-voltage dielectrophoresis-based microfluidic systems: A review.

Dielectrophoretically driven microfluidic devices have demonstrated great applicability in biomedical engineering, diagnostic medicine, and biological research. One of the potential fields of application for this technology is in point-of-care (POC) devices, ideally allowing for portable, fully integrated, easy to use, low-cost diagnostic platforms. Two main approaches exist to induce dielectrophoresis (DEP) on suspended particles, that is, electrode-based DEP and insulator-based DEP, each featuring different advantages and disadvantages. However, a shared concern lies in the input voltage used to generate the electric field necessary for DEP to take place. Therefore, input voltage can determine portability of a microfluidic device. This review outlines the recent advances in reducing stimulation voltage requirements in DEP-driven microfluidics.

Simultaneous Determination of Linear and Nonlinear Electrophoretic Mobilities of Cells and Microparticles.

Direct-current insulator-based electrokinetics (DC-iEK) is a branch of microfluidics that has demonstrated to be an attractive and efficient technique for manipulating micro- and nano- particles, including microorganisms. A unique feature of DC-iEK devices is that nonlinear EK effects are enhanced by the presence of regions of higher field intensity between the insulating structures. Accurate computational models, describing particle and cell behavior, are crucial to optimize the design and improve the performance of DC-iEK devices. The electrokinetic equilibrium condition (EEEC) is a recently introduced fundamental concept that has radically shifted the perspective behind the analysis of particle manipulation in these microfluidic devices. The EEEC takes into consideration previously neglected nonlinear effects on particle migration and indicates that these effects are central to control particle motion in DC-iEK devices. In this study, we present a simultaneous experimental characterization of linear and nonlinear electrokinetic (EK) parameters, that is, the electrophoretic mobility (µEP(1)), the particle zeta potential (ζP), the EEEC, and the electrophoretic mobility of the second kind (µEP(3)), for four types of polystyrene microparticles and four cell strains. For this, we studied the electromigration of polystyrene microparticles ranging in size from 2 to 6.8 µm, three bacteria strains (B. cereus, E. coli, and S. enterica) and a yeast cell (S. cerevisiae), ranging in size from 1 to 6.3 µm, in a polydimethylsiloxane (PDMS) microfluidic channel with a rectangular cross-section. The results illustrated that electrokinetic particle trapping can occur by linear and nonlinear electrophoresis and electroosmosis reaching an equilibrium, without the presence of insulating posts. The experimentally measured parameters reported herein will allow optimizing the design of future DC-iEK devices for a wide range of applications (e.g., to separate multiple kinds of particles and microorganisms) and for developing computational models that better represent reality.

Direct Current Electrokinetic Particle Trapping in Insulator-Based Microfluidics: Theory and Experiments.

The classic theory of direct-current (DC) insulator-based dielectrophoresis (iDEP) considers that, in order to elicit particle trapping, dielectrophoretic (DEP) velocity counterbalances electrokinetic (EK) motion, that is, electrophoresis (EP) and electro-osmotic flow (EOF). However, the particle velocity DEP component requires empirical correction factors (sometimes as high as 600) to account for experimental observations, suggesting the need for a refined model. Here, we show that, when applied to particle suspensions, a high-magnitude DC uniform electric field induces nonlinear particle velocities, leading to particle flow reversal beyond a critical field magnitude, referred to as the EK equilibrium condition. We further demonstrate that this particle motion can be described through an exploratory induced-charge EP nonlinear model. The model predictions were validated under an insulator-based microfluidic platform demonstrating predictive particle trapping for three different particle sizes (with an estimation error < 10%, not using correction factors). Our findings suggest that particle motion and trapping in "DC-iDEP" devices are dominated by EP and EOF, rather than by DEP effects.

Electrokinetically Driven Exosome Separation and Concentration Using Dielectrophoretic-Enhanced PDMS-Based Microfluidics.

Exosomes are a specific subpopulation of extracellular vesicles that have gained interest because of their many potential biomedical applications. However, exosome isolation and characterization are the first steps toward designing novel applications. This work presents a direct current-insulator-based dielectrophoretic (DC-iDEP) approach to simultaneously capture and separate exosomes by size. To do so, a microdevice consisting of a channel with two electrically insulating post sections was designed. Each section was tailored to generate different nonuniform spatial distributions of the electric field and, therefore, different dielectrophoretic forces acting on exosomes suspended in solution. Side channels were placed adjacent to each section to allow sample recovery. By applying an electric potential difference of 2000 V across the length of the main channel, dielectrophoretic size-based separation of exosomes was observed in the device. Analysis of particle size in each recovered fraction served to assess exosome separation efficiency. These findings show that iDEP can represent a first step toward designing a high-throughput, fast, and robust microdevice capable of capturing and discriminating different subpopulations of exosomes based on their size.

Recent advances and challenges in the recovery and purification of cellular exosomes.

Exosomes are nanovesicles secreted by most cellular types that carry important biochemical compounds throughout the body with different purposes, playing a preponderant role in cellular communication. Because of their structure, physicochemical properties and stability, recent studies are focusing in their use as nanocarriers for different therapeutic compounds for the treatment of different diseases ranging from cancer to Parkinson's disease. However, current bioseparation protocols and methodologies are selected based on the final exosome application or intended use and present both advantages and disadvantages when compared among them. In this context, this review aims to present the most important technologies available for exosome isolation while discussing their advantages and disadvantages and the possibilities of being combined with other strategies. This is critical since the development of novel exosome-based therapeutic strategies will be constrained to the effectiveness and yield of the selected downstream purification methodologies for which a thorough understanding of the available technological resources is needed.

Joule heating effects in optimized insulator-based dielectrophoretic devices: An interplay between post geometry and temperature rise.

Insulator-based dielectrophoresis (iDEP) is the electrokinetic migration of polarized particles when subjected to a non-uniform electric field generated by the inclusion of insulating structures between two remote electrodes. Electrode spacing is considerable in iDEP systems when compared to electrode-based DEP systems, therefore, iDEP systems require high voltages to achieve efficient particle manipulation. A consequence of this is the temperature increase within the channel due to Joule heating effects, which, in some cases, can be detrimental when manipulating biological samples. This work presents an experimental and modeling study on the increase in temperature inside iDEP devices. For this, we studied seven distinct channel designs that mainly differ from each other in their post array characteristics: post shape, post size and spacing between posts. Experimental results obtained using a custom-built copper Resistance Temperature Detector, based on resistance changes, show that the influence of the insulators produces a difference in temperature rise of approximately 4°C between the designs studied. Furthermore, a 3D COMSOL model is also introduced to evaluate heat generation and dissipation, which is in good agreement with the experiments. The model allowed relating the difference in average temperature for the geometries under study to the electric resistance posed by the post array in each design.

Simple Approach to Reducing Particle Trapping Voltage in Insulator-Based Dielectrophoretic Systems.

Insulator-based dielectrophoresis (iDEP) is a microfluidic technique used for particle analysis in a wide array of applications. Significant efforts are dedicated to improve iDEP systems by reducing voltage requirements. This study assesses how the performance of an iDEP system, in terms of particle trapping, depends on the number of insulating obstacles longitudinally present in the microchannel. In analogy with Kirchhoff's loop rule, iDEP systems were analyzed as a series combination of electrical resistances, where the equivalent resistance of the post array is composed by a number of individual resistors (columns of insulating posts). It was predicted by the COMSOL model, and later confirmed by experimental results, that reducing the number of columns of insulating posts significantly affects the electric field distribution, decreasing the required voltage to dielectrophoretically trap particles within the post array. As an application, it was demonstrated that decreasing the number of columns in the post array allows for the dielectrophoretic trapping of nanometer-scale particles at voltages well below those reported in previous similar iDEP systems. These findings illustrate how the iDEP channel configuration can be customized for specific applications.