Optically induced electrothermal microfluidic tweezers in bio-relevant media.

Non-contact micro-manipulation tools have enabled invasion-free studies of fragile synthetic particles and biological cells. Rapid electrokinetic patterning (REP) traps target particles/cells, suspended in an electrolyte, on an electrode surface. This entrapment is electrokinetic in nature and thus depends strongly on the suspension medium's properties. REP has been well characterized for manipulating synthetic particles suspended in low concentration salt solutions (~ 2 mS/m). However, it is not studied as extensively for manipulating biological cells, which introduces an additional level of complexity due to their limited viability in hypotonic media. In this work, we discuss challenges posed by isotonic electrolytes and suggest solutions to enable REP manipulation in bio-relevant media. Various formulations of isotonic media (salt and sugar-based) are tested for their compatibility with REP. REP manipulation is observed in low concentration salt-based media such as 0.1× phosphate buffered saline (PBS) when the device electrodes are passivated with a dielectric layer. We also show manipulation of murine pancreatic cancer cells suspended in a sugar-based (8.5% w/v sucrose and 0.3% w/v dextrose) isotonic medium. The ability to trap mammalian cells and deposit them in custom patterns enables high-impact applications such as determining their biomechanical properties and 3D bioprinting for tissue scaffolding.

Particle-Induced Electrostatic Repulsion within an Electric Curtain Operating below the Paschen Limit.

The electric curtain is a platform developed to lift and transport charged particles in air. Its premise is the manipulation of charged particles; however, fewer investigations isolate dielectric forces that are observed at lower voltages (i.e., less than the Paschen limit). This work focuses on observations of simultaneous dielectrophoretic and electrostatic forces. The electric curtain was a printed circuit board with interdigitated electrodes (0.020 inch width and spacing) coated with a layer of polypropylene, where a standing wave or travelling wave AC signal was applied (50 Hz) to produce an electric field below the Paschen limit. Soda lime glass beads (180-212 µm) demonstrated oscillatory rolling via dielectrophoretic forces. In addition, several particles simultaneously experienced rapid projectile repulsion, a behavior consistent with electrostatic phenomena. This second result is discussed as a particle-induced local increase in the electric field, with simulations demonstrating that a particle in close proximity to the curtain's surface produces a local field enhancement of over 2.5 times. The significance of this is that individual particles themselves can trigger electrostatic repulsion in an otherwise dielectric system. These results could be used for advanced applications where particles themselves provided triggered responses, perhaps for selective sorting of micrometer particles in air.

Coherently tunable metalens tweezers for optofluidic particle routing.

Nanophotonic particle manipulation exploits unique light shaping capabilities of nanophotonic devices to trap, guide, rotate and propel particles in microfluidic channels. Recent introduction of metalens into microfluidics research demonstrates the new capability of using nanophotonics devices for far-field optical manipulation. In this work we demonstrate, via numerical simulation, the first tunable metalens tweezers that function under dual-beam illumination. The phase profile of the metalens is modulated by controlling the relative strength and phase of the two coherent incident light beams. As a result, the metalens creates a thin sheet of focus inside a microchannel. Changes to the illumination condition allow the focus to be swept across the microchannel, thereby producing a controllable and reconfigurable path for particle transport. Particle routing in a Y-branch junction, for both nano- and microparticles, is evaluated as an example functionality for the tunable metalens tweezers. This work shows that tunable far-field particle manipulation can be achieved using near-field nano-engineering and coherent control, opening a new way for the integration of nanophotonics and microfluidics.

Scaling law analysis of electrohydrodynamics and dielectrophoresis for isomotive dielectrophoresis microfluidic devices.

Isomotive dielectrophoresis (isoDEP) is a unique DEP geometrical configuration where the gradient of the field-squared ( ∇Erms2 ) is constant. IsoDEP analyzes polarizable particles based on their magnitude and direction of translation. Particle translation is a function of the polarizability of both the particles and suspending medium, the particles' size and shape, and the frequency of the electric field. However, other electrokinetics act on the particles simultaneously, including electrothermal hydrodynamics. Hence, to maximize the DEP force relative to over electrokinetic forces, design parameters such as microchannel geometry, fabrication materials, and applied electric field must be properly tuned. In this work, scaling law analyses were developed to derive design rules, relative to particle diameter, to reduce unwanted electrothermal hydrodynamics relative to DEP-induced particle translation. For a particle suspended in 10 mS/m media, if the channel width and height are below ten particle diameters, the electrothermal-driven flow is reduced by ∼500 times compared to a channel that is 250 particles diameters in width and height. Replacing glass with silicon as the device's underlying substrate for an insulative-based isoDEP reduces the electrothermal induced flow approximately 20 times less.

Image-based sorting and negative dielectrophoresis for high purity cell and particle separation.

Microelectrode arrays are used to sort single fluorescently labeled cells and particles as they flow through a microfluidic channel using dielectrophoresis. Negative dielectrophoresis is used to create a "Dielectrophoretic virtual channel" that runs along the center of the microfluidic channel. By switching the polarity of the electrodes, the virtual channel can be dynamically reconfigured to direct particles along a different path. This is demonstrated by sorting particles into two microfluidic outlets, controlled by an automated system that interprets video data from a color camera and makes complex sorting decisions based on color, intensity, size, and shape. This enables the rejection of particle aggregates and other impurities, and the system is optimized to isolate high purity populations from a heterogeneous sample. Green beads are isolated from an excess of red beads with 100% purity at a rate of up to 0.9 particles per second, in addition application to the sorting of osteosarcoma and human bone marrow cells is evidenced. The extension of Dielectrophoretic Virtual Channels to an arbitrary number of sorting outputs is examined, with design, simulation, and experimental verification of two alternate geometries presented and compared.

A novel portable filtration system for sampling and concentration of microorganisms: Demonstration on marine microalgae with subsequent quantification using IC-NASBA.

This paper presents a novel portable sample filtration/concentration system, designed for use on samples of microorganisms with very low cell concentrations and large volumes, such as water-borne parasites, pathogens associated with faecal matter, or toxic phytoplankton. The example application used for demonstration was the in-field collection and concentration of microalgae from seawater samples. This type of organism is responsible for Harmful Algal Blooms (HABs), an example of which is commonly referred to as "red tides", which are typically the result of rapid proliferation and high biomass accumulation of harmful microalgal species in the water column or at the sea surface. For instance, Karenia brevis red tides are the cause of aquatic organism mortality and persistent blooms may cause widespread die-offs of populations of other organisms including vertebrates. In order to respond to, and adequately manage HABs, monitoring of toxic microalgae is required and large-volume sample concentrators would be a useful tool for in situ monitoring of HABs. The filtering system presented in this work enables consistent sample collection and concentration from 1 L to 1 mL in five minutes, allowing for subsequent benchtop sample extraction and analysis using molecular methods such as NASBA and IC-NASBA. The microalga Tetraselmis suecica was successfully detected at concentrations ranging from 2 × 105 cells/L to 20 cells/L. Karenia brevis was also detected and quantified at concentrations between 10 cells/L and 106 cells/L. Further analysis showed that the filter system, which concentrates cells from very large volumes with consequently more reliable sampling, produced samples that were more consistent than the independent non-filtered samples (benchtop controls), with a logarithmic dependency on increasing cell numbers. This filtering system provides simple, rapid, and consistent sample collection and concentration for further analysis, and could be applied to a wide range of different samples and target organisms in situations lacking laboratories.

Field-effect sensors - from pH sensing to biosensing: sensitivity enhancement using streptavidin-biotin as a model system.

Field-Effect Transistor sensors (FET-sensors) have been receiving increasing attention for biomolecular sensing over the last two decades due to their potential for ultra-high sensitivity sensing, label-free operation, cost reduction and miniaturisation. Whilst the commercial application of FET-sensors in pH sensing has been realised, their commercial application in biomolecular sensing (termed BioFETs) is hindered by poor understanding of how to optimise device design for highly reproducible operation and high sensitivity. In part, these problems stem from the highly interdisciplinary nature of the problems encountered in this field, in which knowledge of biomolecular-binding kinetics, surface chemistry, electrical double layer physics and electrical engineering is required. In this work, a quantitative analysis and critical review has been performed comparing literature FET-sensor data for pH-sensing with data for sensing of biomolecular streptavidin binding to surface-bound biotin systems. The aim is to provide the first systematic, quantitative comparison of BioFET results for a single biomolecular analyte, specifically streptavidin, which is the most commonly used model protein in biosensing experiments, and often used as an initial proof-of-concept for new biosensor designs. This novel quantitative and comparative analysis of the surface potential behaviour of a range of devices demonstrated a strong contrast between the trends observed in pH-sensing and those in biomolecule-sensing. Potential explanations are discussed in detail and surface-chemistry optimisation is shown to be a vital component in sensitivity-enhancement. Factors which can influence the response, yet which have not always been fully appreciated, are explored and practical suggestions are provided on how to improve experimental design.

Electrothermal pumping with interdigitated electrodes and resistive heaters.

Interdigitated electrodes are used in electrokinetic lab-on-a-chip devices for dielectrophoretic trapping and characterization of suspended particles, as well as the production of field-induced fluid flow via AC electroosomosis and electrothermal mechanisms. However, the optimum design for dielectrophoresis, that if symmetrical electrodes, cannot induce bulk electrohydrodynamic pumping. In addition, the mechanism of intrinsic electrothermal pumping is affected by the properties of the fluid, with thermal fields being generated by Joule Heating. This work demonstrates the incorporation of an underlying thin film heater, electrically isolated from the interdigitated electrodes by an insulator layer, to enhance bulk electrothermal pumping. The use of integrated heaters allows the thermal field generation to be controlled independently of the electric field. Numerical simulations are performed to demonstrate the importance of geometrical arrangement of the heater with respect to the interdigitated electrodes, as well as electrode size, spacing, and arrangement. The optimization of such a system is a careful balance between electrokinetics, heat transfer, and fluid dynamics. The heater location and electrode spacing influence the rate of electrothermal pumping significantly more than electrode width and insulator layer thickness. This demonstration will aid in the development of microfluidic electrokinetic systems that want to utilize the advantages associated with electrothermal pumping while simultaneously applying other lab-on-a-chip electrokinetics like dielectrophoresis.

Acid-base dissociation mechanisms and energetics at the silica-water interface: An activationless process.

HYPOTHESIS: Silanol groups at the silica-water interface determine not only the surface charge, but also have an important role in the binding of ions and biomolecules. As the pH is increased above pH 2, the silica surface develops a net negative charge primarily due to deprotonation of the silanol group. An improved understanding of the energetics and mechanisms of this fundamentally important process would further understanding of the relevant dynamics. SIMULATIONS: Density Functional Theory ab initio molecular dynamics and geometry optimisations were used to investigate the mechanisms of surface neutralisation and charging in the presence of OH(-) and H3O(+) respectively. This charging mechanism has received little attention in the literature. FINDINGS: The protonation or deprotonation of isolated silanols in the presence of H3O(+) or OH(-), respectively, was shown to be a highly rapid, exothermic reaction with no significant activation energy. This process occurred via a concerted motion of the protons through 'water wires'. Geometry optimisations of large water clusters at the silica surface demonstrated proton transfer to the surface occurring via the rarely discussed 'proton holes' mechanism. This indicates that surface protonation is possible even when the hydronium ion is distant (at least 4 water molecules separation) from the surface.

An optoelectrokinetic technique for programmable particle manipulation and bead-based biosignal enhancement.

Technologies that can enable concentration of low-abundance biomarkers are essential for early diagnosis of diseases. In this study, an optoelectrokinetic technique, termed Rapid Electrokinetic Patterning (REP), was used to enable dynamic particle manipulation in bead-based bioassays. Various manipulation capabilities, such as micro/nanoparticle aggregation, translation, sorting and patterning, were developed. The technique allows for versatile multi-parameter (voltage, light intensity and frequency) based modulation and dynamically addressable manipulation with simple device fabrication. Signal enhancement of a bead-based bioassay was demonstrated using dilute biotin-fluorescein isothiocyanate (FITC) solutions mixed with streptavidin-conjugated particles and rapidly concentrated with the technique. As compared with a conventional ELISA reader, the REP-enabled detection achieved a minimal readout of 3.87 nM, which was a 100-fold improvement in sensitivity. The multi-functional platform provides an effective measure to enhance detection levels in more bead-based bioassays.

Higher-order dielectrophoresis of nonspherical particles.

Higher-order terms of dielectrophoretic (DEP) force are commonly ignored by invoking the simplifying dipole approximation. Concurrently, the trend towards micro- and nano-electrode structures in DEP design is bringing about an increasing number of instances where the approximation is expected to lose reliability. The case is severe for nonspherical particles (the shape of many biological particles) due to the shape-dependent nature of dielectric polarization. However, there is a lack of analytical means to determine multipole moments of nonspherical particles, numerical calculations of the same are regarded as unreliable, and there is a prevalence for higher-order force considerations to be ignored. As a result, the dipole approximation is used and/or nonspherical particles are approximated as spheres. This work proves the inefficacy of current qualitative criteria for the reliability of the dipole approximation and presents a quantitative substitute, with verified accuracy, that enables precise determination of the extent to which the dipole approximation would be reliable, and if found unreliable, corrects the approximation by adding second- and third-order terms of the DEP force. The effects of field nonuniformity, electrode design, and particle shape and aspect ratio on the significance of higher-order DEP forces is quantitatively analyzed. The results show that higher-order DEP forces are indeed of substantially increased significance for nonspherical particles; in the cases examined in this work, multipolar terms are seen to constitute more than 40% of the total force on ellipsoidal and cylindrical particles. It is further shown that approximating nonspherical particles as spheres of similar dimensions is subject to substantial error. Last, the substantial importance of the electrode design in influencing higher-order forces is shown.

Continuous separation of colloidal particles using dielectrophoresis.

Dielectrophoresis is the movement of particles in nonuniform electric fields and has been of interest for application to manipulation and separation at and below the microscale. This technique has the advantages of being noninvasive, nondestructive, and noncontact, with the movement of particle achieved by means of electric fields generated by miniaturized electrodes and microfluidic systems. Although the majority of applications have been above the microscale, there is increasing interest in application to colloidal particles around a micron and smaller. This paper begins with a review of colloidal and nanoscale dielectrophoresis with specific attention paid to separation applications. An innovative design of integrated microelectrode array and its application to flow-through, continuous separation of colloidal particles is then presented. The details of the angled chevron microelectrode array and the test microfluidic system are then discussed. The variation in device operation with applied signal voltage is presented and discussed in terms of separation efficiency, demonstrating 99.9% separation of a mixture of colloidal latex spheres.

Hybrid opto-electric manipulation in microfluidics-opportunities and challenges.

Hybrid opto-electric manipulation in microfluidics/nanofluidics refers to a set of methodologies employing optical modulation of electrokinetic schemes to achieve particle or fluid manipulation at the micro- and nano-scale. Over the last decade, a set of methodologies, which differ in their modulation strategy and/or the length scale of operation, have emerged. These techniques offer new opportunities with their dynamic nature, and their ability for parallel operation has created novel applications and devices. Hybrid opto-electric techniques have been utilized to manipulate objects ranging in diversity from millimetre-sized droplets to nano-particles. This review article discusses the underlying principles, applications and future perspectives of various hybrid opto-electric techniques that have emerged over the last decade under a unified umbrella.

Integrated systems for rapid point of care (PoC) blood cell analysis.

Counting the different subpopulations of cells in a fingerprick of human blood is important for a number of clinical point-of-care (PoC) applications. It is a challenge to demonstrate the integration of sample preparation and detection techniques in a single platform. In this paper we demonstrate a generic microfluidic platform that combines sample processing and characterisation and enumeration in a single, integrated system. Results of microfluidic 3-part differential leukocyte (granulocyte, lymphocyte and monocyte) counts, together with erythrocyte and thrombocyte (platelet) counts, in human blood are shown and corroborated with results from hospital clinical laboratory analysis.

Optically modulated electrokinetic manipulation and concentration of colloidal particles near an electrode surface.

We study a recently demonstrated AC electrokinetic technique for manipulation and concentration of colloidal particles on an electrode surface. The technique uses indium tin oxide (ITO)-based parallel-plate electrodes on which highly localized infrared (1064 nm) laser illumination is shone. We show that the highly localized laser illumination leads to a highly nonuniform heating of the electrode substrate, which in turn drives an electrothermal microvortex resulting in a rapid transport of particles toward the illuminated site. Hundreds of polystyrene particles, with diameters ranging from 2.0 to 0.1 microm, suspended in a low conductivity solution (2.0 mS/m) could be aggregated at selected locations on the electrode by activating the laser illumination at suitable AC frequencies. Subsequent deactivation of the laser illumination causes the particles to scatter, and we explore this dynamical behavior for 1.0 microm particles using Delaunay tessellations and high-speed videography. We establish that drag from the electrothermal microvortex acts against a repulsive force, which decreases with increasing AC frequency, to create stable particle clusters. Moreover, experimentally we show that this particle capturing technique can be characterized by a critical frequency: a frequency at which the captured colloidal particle cluster becomes unstable and particles are carried away into the bulk by the electrothermal microvortex. This critical frequency increases with decreasing particle diameter for similar particles. For 0.1 microm particles, comparison of aggregation at different AC frequencies is achieved by the comparison of fluorescent intensity profiles of the aggregations.

Negative DEP traps for single cell immobilisation.

We present a novel design of micron-sized particle trap that uses negative dielectrophoresis (nDEP) to trap cells in high conductivity physiological media. The design is scalable and suitable for trapping large numbers of single cells. Each trap has one electrical connection and the design can be extended to produce a large array. The trap consists of a metal ring electrode and a surrounding ground plane, which create a closed electric field cage in the centre. The operation of the device was demonstrated by trapping single latex spheres and HeLa cells against a moving fluid. The dielectrophoretic holding force was determined experimentally by measuring the displacement of a trapped particle in a moving fluid. This was then compared with theory by numerically solving the electric field for the electrodes and calculating the trapping force, demonstrating good agreement. Analysis of the 80 microm diameter trap showed that a 15.6 microm diameter latex particle could be held with a force of 23 pN at an applied voltage of 5 V peak-peak.

A simple, optically induced electrokinetic method to concentrate and pattern nanoparticles.

We demonstrate an optically induced electrokinetic technique that continuously concentrates nanoparticles on the surface of a parallel plate electrode that is biased with an AC signal. A highly focused beam of near-infrared light (1064 nm) was applied, inducing an electrothermal microfluidic vortex that carried nanoparticles to its center where they were accumulated. This technique was demonstrated with 49 nm and 100 nm fluorescent polystyrene particles and characterized as a function of applied AC frequency and voltage. With this technique the location and shape of colloidal concentration was reconfigured by controlling the optical landscape, yielding dynamic control of the aggregation. Colloidal concentration was demonstrated with a plain parallel plate electrode configuration without the need of photoconductive materials or complex microfabrication procedures.

Formation of artificial lipid bilayers using droplet dielectrophoresis.

We describe the formation of artificial bilayer lipid membranes (BLMs) by the controlled, electrical manipulation of aqueous droplets immersed in a lipid-alkane solution. Droplet movement was generated using dielectrophoresis on planar microelectrodes covered in a thin insulator. Droplets, surrounded by lipid monolayers, were brought into contact and spontaneously formed a BLM. The method produced BLMs suitable for single-channel recording of membrane protein activity and the technique can be extended to create programmable BLM arrays and networks.

Analytical solutions of ac electrokinetics in interdigitated electrode arrays: electric field, dielectrophoretic and traveling-wave dielectrophoretic forces.

Analysis of the movement of particles in a nonuniform field requires accurate knowledge of the electric field distribution in the system. This paper describes a method for analytically solving the electric field distribution above interdigitated electrode arrays used for dielectrophoresis (DEP) and traveling wave dielectrophoresis (twDEP), using the Schwarz-Christoffel mapping method. The electric field solutions are used to calculate the dielectrophoretic force in both cases, and the traveling wave dielectrophoretic force and the electrorotational torque for the twDEP case. This method requires no approximations and can take into account the Neumann boundary condition used to represent an insulating lid and lower substrate. The analytical results of the electric field distributions are validated for different geometries by comparison with numerical simulations using the finite element method.

High speed multi-frequency impedance analysis of single particles in a microfluidic cytometer using maximum length sequences.

A novel impedance spectroscopy technique has been developed for high speed single biological particle analysis. A microfluidic cytometer is used to measure the impedance of single micrometre sized latex particles at high speed across a range of frequencies. The setup uses a technique based on maximum length sequence (MLS) analysis, where the time-dependent response of the system is measured in the time domain and transformed into the impulse response using fast M-sequence transform (FMT). Finally fast Fourier transform (FFT) is applied to the impulse response to give the transfer-function of the system in the frequency domain. It is demonstrated that the MLS technique can give multi-frequency (broad-band) measurement in a short time period (ms). The impedance spectra of polystyrene micro-beads are measured at 512 evenly distributed frequencies over a range from 976.5625 Hz to 500 kHz. The spectral information for each bead is obtained in approximately 1 ms. Good agreement is shown between the MLS data and both circuit simulations and conventional AC single frequency measurements.