Ultrafast microfluidics using surface acoustic waves

We demonstrate that surface acoustic waves (SAWs), nanometer amplitude Rayleigh waves driven at megahertz order frequencies propagating on the surface of a piezoelectric substrate, offer a powerful method for driving a host of extremely fast microfluidic actuation and micro∕bioparticle manipulation schemes. We show that sessile drops can be translated rapidly on planar substrates or fluid can be pumped through microchannels at 1–10 cm∕s velocities, which are typically one to two orders quicker than that afforded by current microfluidic technologies. Through symmetry-breaking, azimuthal recirculation can be induced within the drop to drive strong inertial microcentrifugation for micromixing and particle concentration or separation. Similar micromixing strategies can be induced in the same microchannel in which fluid is pumped with the SAW by merely changing the SAW frequency to rapidly switch the uniform through-flow into a chaotic oscillatory flow by exploiting superpositioning of the irradiated sound waves from the sidewalls of the microchannel. If the flow is sufficiently quiescent, the nodes of the transverse standing wave that arises across the microchannel also allow for particle aggregation, and hence, sorting on nodal lines. In addition, the SAW also facilitates other microfluidic capabilities. For example, capillary waves excited at the free surface of a sessile drop by the SAW underneath it can be exploited for micro∕nanoparticle collection and sorting at nodal points or lines at low powers. At higher powers, the large accelerations off the substrate surface as the SAW propagates across drives rapid destabilization of the drop free surface giving rise to inertial liquid jets that persist over 1–2 cm in length or atomization of the entire drop to produce 1–10 μm monodispersed aerosol droplets, which can be exploited for ink-jet printing, mass spectrometry interfacing, or pulmonary drug delivery. The atomization of polymer∕protein solutions can also be used for the rapid synthesis of 150–200 nm polymer∕protein particles or biodegradable polymeric shells in which proteins, peptides, and other therapeutic molecules are encapsulated within for controlled release drug delivery. The atomization of thin films behind a translating drop containing polymer solutions also gives rise to long-range spatial ordering of regular polymer spots whose size and spacing are dependent on the SAW frequency, thus offering a simple and powerful method for polymer patterning without requiring surface treatment or physical∕chemical templating.     

Spatial concentration distribution analysis of cells in electrode-multilayered microchannel by dielectric property measurement

The spatial concentration distribution of cells in a microchannel is measured by combining the dielectric properties of cells with the specific structure of the electrode-multilayered microchannel. The dielectric properties of cells obtained with the impedance spectroscopy method includes the cell permittivity and dielectric relaxation, which corresponds to the cell concentration and structure. The electrode-multilayered microchannel is constructed by 5 cross-sections, and each cross-section contains 5 electrode-layers embedded with 16 micro electrodes. In the experiment, the dielectric properties of cell suspensions with different volume concentrations are measured with different electrode-combinations corresponding to different electric field distributions. The dielectric relaxations of different cell concentrations are compared and discussed with the Maxwell-Wagner dispersion theory, and the relaxation frequencies are analysed by a cell polarization model established based on the Hanai cell model. Moreover, a significant linear relationship with AC frequency dependency between relative permittivity and cell concentration was found, which provides a promising way to on-line estimate cell concentration in microchannel. Finally, cell distribution in 1 cross-section of the microchannel (X and Y directions) was measured with different electrode-combinations using the dielectric properties of cell suspensions, and cell concentration distribution along the microchannel (Z direction) was visualized at flowing state. The present cell spatial sensing study provides a new approach for 3 dimensional non-invasive online cell sensing for biological industry.     

Development of assembled microchannel resonator as an alternative fabrication method of a microchannel resonator for mass sensing in flowing liquid

We propose an alternate fabrication technique of microchannel resonators based on an assembly method of three separate parts to form a microchannel resonator on a chip. The capability of the assembled microchannel resonator to detect mass is confirmed by injecting two liquids with different densities. The experimental and theoretical values of the resonator frequency shift are in agreement with each other, which confirms the consistency of the device. The noise level of the device is estimated from the Allan variance plot, so the minimum detectable mass of 230 fg after 16 s of operation is expected. By considering the time of the practical application of 1 ms, it is found that a detectable mass of around 8.51 pg is estimated, which is applicable for detecting flowing microparticles. The sub-pico to a few picogram levels of detection will be applicable for the mass analysis of flowing microparticles such as single cells and will be greatly beneficial for many fields such as chemistry, medicine, biology, and single-cell analysis.     

Three-dimensional diamagnetic particle deflection in ferrofluid microchannel flows

Magnetic field-induced particle manipulation is a promising technique for biomicrofluidics applications. It is simple, cheap, and also free of fluid heating issues that accompany other common electric, acoustic, and optical methods. This work presents a fundamental study of diamagnetic particle motion in ferrofluid flows through a rectangular microchannel with a nearby permanent magnet. Due to their negligible magnetization relative to the ferrofluid, diamagnetic particles experience negative magnetophoresis and are repelled away from the magnet. The result is a three-dimensionally focused particle stream flowing near the bottom outer corner of the microchannel that is the farthest to the center of the magnet and hence has the smallest magnetic field. The effects of the particle's relative position to the magnet, particle size, ferrofluid flow rate, and concentration on this three-dimensional diamagnetic particle deflection are systematically studied. The obtained experimental results agree quantitatively with the predictions of a three-dimensional analytical model.     

Transport of particles and microorganisms in microfluidic channels using rectified ac electro-osmotic flow

A new method is demonstrated to transport particles, cells, and other microorganisms using rectified ac electro-osmotic flows in open microchannels. The rectified flow is obtained by synchronous zeta potential modulation with the driving potential in the microchannel. Experiments were conducted to transport both neutral, charged particles, and microorganisms of various sizes. A maximum speed of 50 μm∕s was obtained for 8 μm polystyrene beads, without any electrolysis, using a symmetrical square waveform driving electric field of 5 V∕mm at 10 Hz and a 360 V gate potential with its polarity synchronized with the driving potential (phase lag=0°).     

Cell separation and transportation between two miscible fluid streams using ultrasound

This paper presents a two-stream microfluidic system for transporting cells or micro-sized particles from one fluid stream to another by acoustophoresis. The two fluid streams, one being the original suspension and the other being the destination fluid, flow parallel to each other in a microchannel. Using a half-wave acoustic standing wave across the channel width, cells or particles with positive acoustic contrast factors are moved to the destination fluid where the pressure nodal line lies. By controlling the relative flow rate of the two fluid streams, the pressure nodal line can be maintained at a specific offset from the fluid interface within the destination fluid. Using this transportation method, particles or cells of different sizes and mechanical properties can be separated. The cells experiencing a larger acoustic radiation force are separated and transported from the original suspension to the destination fluid stream. The other particles or cells experiencing a smaller acoustic radiation force continue flowing in the original solution. Experiments were conducted to demonstrate the effective separation of polystyrene microbeads of different sizes (3 μm and 10 μm) and waterborne parasites (Giardia lamblia and Cryptosporidium parvum). Diffusion occurs between the two miscible fluids, but it was found to have little effects on the transport and separation process, even when the two fluids have different density and speed of sound.     

Elasto-inertial particle focusing under the viscoelastic flow of DNA solution in a square channel

Particle focusing is an essential step in a wide range of applications such as cell counting and sorting. Recently, viscoelastic particle focusing, which exploits the spatially non-uniform viscoelastic properties of a polymer solution under Poiseuille flow, has attracted much attention because the particles are focused along the channel centerline without any external force. Lateral particle migration in polymer solutions in square channels has been studied due to its practical importance in lab-on-a-chip applications. However, there are still many questions about how the rheological properties of the medium alter the equilibrium particle positions and about the flow rate ranges for particle focusing. In this study, we investigated lateral particle migration in a viscoelastic flow of DNA solution in a square microchannel. The elastic property is relevant due to the long relaxation time of a DNA molecule, even when the DNA concentration is extremely low. Further, the shear viscosity of the solution is essentially constant irrespective of shear rate. Our current results demonstrate that the particles migrate toward the channel centerline and the four corners of a square channel in the dilute DNA solution when the inertia is negligible (elasticity-dominant flow). As the flow rate increases, the multiple equilibrium particle positions are reduced to a single file along the channel centerline, due to the elasto-inertial particle focusing mechanism. The current results support that elasto-inertial particle focusing mechanism is a universal phenomenon in a viscoelastic fluid with constant shear viscosity (Boger fluid). Also, the effective flow rate ranges for three-dimensional particle focusing in the DNA solution were significantly higher and wider than those for the previous synthetic polymer solution case, which facilitates high throughput analysis of particulate systems. In addition, we demonstrated that the DNA solution can be applied to focus a wide range of particle sizes in a single channel and also align red blood cells without any significant deformation.     

Modulation of rotation-induced lift force for cell filtration in a low aspect ratio microchannel

Cell filtration is a critical step in sample preparation in many bioapplications. Herein, we report on a simple, filter-free, microfluidic platform based on hydrodynamic inertial migration. Our approach builds on the concept of two-stage inertial migration which permits precise prediction of microparticle position within the microchannel. Our design manipulates equilibrium positions of larger microparticles by modulating rotation-induced lift force in a low aspect ratio microchannel. Here, we demonstrate filtration of microparticles with extreme efficiency (>99%). Using multiple prostate cell lines (LNCaP and human prostate epithelial tumor cells), we show filtration from spiked blood, with 3-fold concentration and >83% viability. Results of a proliferation assay show normal cell division and suggest no negative effects on intrinsic properties. Considering the planar low-aspect-ratio structure and predictable focusing, we envision promising applications and easy integration with existing lab-on-a-chip systems.     

A hydrodynamic focusing microchannel based on micro-weir shear lift force

A novel microflow cytometer is proposed in which the particles are focused in the horizontal and vertical directions by means of the Saffman shear lift force generated within a micro-weir microchannel. The proposed device is fabricated on stress-relieved glass substrates and is characterized both numerically and experimentally using fluorescent particles with diameters of 5 μm and 10 μm, respectively. The numerical results show that the micro-weir structures confine the particle stream to the center of the microchannel without the need for a shear flow. Moreover, the experimental results show that the particles emerging from the micro-weir microchannel pass through the detection region in a one-by-one fashion. The focusing effect of the micro-weir microchannel is quantified by computing the normalized variance of the optical detection signal intensity. It is shown that the focusing performance of the micro-weir structure is equal to 99.76% and 99.57% for the 5-μm and 10-μm beads, respectively. Overall, the results presented in this study confirm that the proposed microcytometer enables the reliable sorting and counting of particles with different diameters.     

Inertial microfluidics in contraction–expansion microchannels: A review

Inertial microfluidics has brought enormous changes in the conventional cell/particle detection process and now become the main trend of sample pretreatment with outstanding throughput, low cost, and simple control method. However, inertial microfluidics in a straight microchannel is not enough to provide high efficiency and satisfying performance for cell/particle separation. A contraction–expansion microchannel is a widely used and multifunctional channel pattern involving inertial microfluidics, secondary flow, and the vortex in the chamber. The strengthened inertial microfluidics can help us to focus particles with a shorter channel length and less processing time. Both the vortex in the chamber and the secondary flow in the main channel can trap the target particles or separate particles based on their sizes more precisely. The contraction–expansion microchannels are also capable of combining with a curved, spiral, or serpentine channel to further improve the separation performance. Some recent studies have focused on the viscoelastic fluid that utilizes both elastic forces and inertial forces to separate different size particles precisely with a relatively low flow rate for the vulnerable cells. This article comprehensively reviews various contraction–expansion microchannels with Newtonian and viscoelastic fluids for particle focusing, separation, and microfluid mixing and provides particle manipulation performance data analysis for the contraction–expansion microchannel design.     

Transportation of single cell and microbubbles by phase-shift introduced to standing leaky surface acoustic waves

A microfluidic device was developed to precisely transport a single cell or multiple microbubbles by introducing phase-shifts to a standing leaky surface acoustic wave (SLSAW). The device consists of a polydimethyl-siloxane (PDMS) microchannel and two phase-tunable interdigital transducers (IDTs) for the generation of the relative phase for the pair of surface acoustic waves (SAW) propagating along the opposite directions forming a standing wave. When the SAW contacts the fluid medium inside the microchannel, some of SAW energy is coupled to the fluid and the SAW becomes the leaky surface wave. By modulating the relative phase between two IDTs, the positions of pressure nodes of the SLSAW in the microchannel change linearly resulting in the transportation of a single cell or microbubbles. The results also reveal that there is a good linear relationship between the relative phase and the displacement of a single cell or microbubbles. Furthermore, the single cell and the microbubbles can be transported over a predetermined distance continuously until they reach the targeted locations. This technique has its distinct advantages, such as precise position-manipulation, simple to implement, miniature size, and noninvasive character, which may provide an effective method for the position-manipulation of a single cell and microbubbles in many biological and biomedical applications.     

Microfluidic conformal coating of non-spherical magnetic particles

We present the conformal coating of non-spherical magnetic particles in a co-laminar flow microfluidic system. Whereas in the previous reports spherical particles had been coated with thin films that formed spheres around the particles; in this article, we show the coating of non-spherical particles with coating layers that are approximately uniform in thickness. The novelty of our work is that while liquid-liquid interfacial tension tends to minimize the surface area of interfaces—for example, to form spherical droplets that encapsulate spherical particles—in our experiments, the thin film that coats non-spherical particles has a non-minimal interfacial area. We first make bullet-shaped magnetic microparticles using a stop-flow lithography method that was previously demonstrated. We then suspend the bullet-shaped microparticles in an aqueous solution and flow the particle suspension with a co-flow of a non-aqueous mixture. A magnetic field gradient from a permanent magnet pulls the microparticles in the transverse direction to the fluid flow, until the particles reach the interface between the immiscible fluids. We observe that upon crossing the oil-water interface, the microparticles become coated by a thin film of the aqueous fluid. When we increase the two-fluid interfacial tension by reducing surfactant concentration, we observe that the particles become trapped at the interface, and we use this observation to extract an approximate magnetic susceptibility of the manufactured non-spherical microparticles. Finally, using fluorescence imaging, we confirm the uniformity of the thin film coating along the entire curved surface of the bullet-shaped particles. To the best of our knowledge, this is the first demonstration of conformal coating of non-spherical particles using microfluidics.     

Development of three-dimensional integrated microchannel-electrode system to understand the particles' movement with electrokinetics

An optical transparent 3-D Integrated Microchannel-Electrode System (3-DIMES) has been developed to understand the particles'' movement with electrokinetics in the microchannel. In this system, 40 multilayered electrodes are embedded at the 2 opposite sides along the 5 square cross-sections of the microchannel by using Micro Electro-Mechanical Systems technology in order to achieve the optical transparency at the other 2 opposite sides. The concept of the 3-DIMES is that the particles are driven by electrokinetic forces which are dielectrophoretic force, thermal buoyancy, electrothermal force, and electroosmotic force in a three-dimensional scope by selecting the excitation multilayered electrodes. As a first step to understand the particles'' movement driven by electrokinetic forces in high conductive fluid (phosphate buffer saline (PBS)) with the 3-DIMES, the velocities of particles'' movement with one pair of the electrodes are measured three dimensionally by Particle Image Velocimetry technique in PBS; meanwhile, low conductive fluid (deionized water) is used as a reference. Then, the particles'' movement driven by the electrokinetic forces is discussed theoretically to estimate dominant forces exerting on the particles. Finally, from the theoretical estimation, the particles'' movement mainly results from the dominant forces which are thermal buoyancy and electrothermal force, while the velocity vortex formed at the 2 edges of the electrodes is because of the electroosmotic force. The conclusions suggest that the 3-DIMES with PBS as high conductive fluid helps to understand the three-dimensional advantageous flow structures for cell manipulation in biomedical applications.     

Inertial cell sorting of microparticle-laden flows: An innovative OpenFOAM-based arbitrary Lagrangian–Eulerian numerical approach

The need for cell and particle sorting in human health care and biotechnology applications is undeniable. Inertial microfluidics has proven to be an effective cell and particle sorting technology in many of these applications. Still, only a limited understanding of the underlying physics of particle migration is currently available due to the complex inertial and impact forces arising from particle–particle and particle–wall interactions. Thus, even though it would likely enable significant advances in the field, very few studies have tried to simulate particle-laden flows in inertial microfluidic devices. To address this, this study proposes new codes (solved in OpenFOAM software) that capture all the salient inertial forces, including the four-way coupling between the conveying fluid and the suspended particles traveling a spiral microchannel. Additionally, these simulations are relatively (computationally) inexpensive since the arbitrary Lagrangian–Eulerian formulation allows the fluid elements to be much larger than the particles. In this study, simulations were conducted for two different spiral microchannel cross sections (e.g., rectangular and trapezoidal) for comparison against previously published experimental results. The results indicate good agreement with experiments in terms of (monodisperse) particle focusing positions, and the codes can readily be extended to simulate two different particle types. This new numerical approach is significant because it opens the door to rapid geometric and flow rate optimization in order to improve the efficiency and purity of cell and particle sorting in biotechnology applications.     

Acoustofluidic phase microscopy in a tilted segmentation-free configuration

A low-cost device for registration-free quantitative phase microscopy (QPM) based on the transport of intensity equation of cells in continuous flow is presented. The method uses acoustic focusing to align cells into a single plane where all cells move at a constant speed. The acoustic focusing plane is tilted with respect to the microscope’s focal plane in order to obtain cell images at multiple focal positions. As the cells are displaced at constant speed, phase maps can be generated without the need to segment and register individual objects. The proposed inclined geometry allows for the acquisition of a vertical stack without the need for any moving part, and it enables a cost-effective and robust implementation of QPM. The suitability of the solution for biological imaging is tested on blood samples, demonstrating the ability to recover the phase map of single red blood cells flowing through the microchip.     

Dynamic radial positioning of a hydrodynamically focused particle stream enabled by a three-dimensional microfluidic nozzle

The ability to confine flows and focus particle streams has become an integral component of the design of microfluidic systems for the analysis of a wide range of samples. Presented here is the implementation of a 3D microfluidic nozzle capable of both focusing particles as well as dynamically positioning those particles in selected flow lamina within the downstream analysis channel. Through the independent adjustment of the three sheath inlet flows, the nozzle controlled the size of a focused stream for 6, 10, and 15 μm polystyrene microparticles. Additional flow adjustment allowed the nozzle to dynamically position the focused particle stream to a specific area within the downstream channel. This unique ability provides additional capability and sample flexibility to the system. In order to gain insight into the fluidic behavior of the system, experimental conditions and results were duplicated within 4.75 μm using a COMSOL Multiphysics^® model to elucidate the structure, direction, proportion, and fate of fluid lamina throughout the nozzle region. The COMSOL Multiphysics model showed that the position and distribution of particles upon entering the nozzle have negligible influence over its focusing ability, extending the experimental results into a wider range of particle sizes and system flow rates. These results are promising for the application of this design to allow for a relatively simple, fast, fully fluidically controlled nozzle for selective particle focusing and positioning for further particle analysis and sorting.     

High-throughput inertial particle focusing in a curved microchannel: Insights into the flow-rate regulation mechanism and process model

In this work, we design and fabricate a miniaturized spiral-shaped microchannel device which can be used for high-throughput particle/cell ordering, enrichment, and purification. To probe into the flow rate regulation mechanism, an experimental investigation is carried out on the focusing behaviors of particles with significantly different sizes in this device. A complete picture of the focusing position shifting process is unfolded to clarify the confusing results obtained from flow regimes with different dominant forces in past research. Specifically, with the increase of the flow rate, particles are observed to first move towards the inner wall under the dominant inertial migration, then stabilize at a specific position and finally shift away from the inner wall due to the alternation of the dominant force. Novel phenomena of focusing instability, co-focusing, and focusing position interchange of differently sized particles are also observed and investigated. Based on the obtained experimental data, we develop and validate, for the first time, a five-stage model of the particle focusing process with increasing flow rate for interpreting particle behaviors in terms of the competition between inertial lift and Dean drag forces. These new experimental findings and the proposed process model provide an important supplement to the existing mechanism of inertial particle flow and enable more flexible and precise particle manipulation. Additionally, we examine the focusing behaviors of bioparticles with a polydisperse size distribution to validate the explored mechanisms and thus help realize efficient enrichment and purification of these particles.     

Transmittance tuning by particle chain polarization in electrowetting-driven droplets

A tiny droplet containing nano∕microparticles commonly handled in digital microfluidic lab-on-a-chip is regarded as a micro-optical component with tunable transmittance at programmable positions for the application of micro-opto-fluidic-systems. Cross-scale electric manipulations of droplets on a millimeter scale as well as suspended particles on a micrometer scale are demonstrated by electrowetting-on-dielectric (EWOD) and particle chain polarization, respectively. By applying electric fields at proper frequency ranges, EWOD and polarization can be selectively achieved in designed and fabricated parallel plate devices. At low frequencies, the applied signal generates EWOD to pump suspension droplets. The evenly dispersed particles reflect and∕or absorb the incident light to exhibit a reflective or dark droplet. When sufficiently high frequencies are used on to the nonsegmented parallel electrodes, a uniform electric field is established across the liquid to polarize the dispersed neutral particles. The induced dipole moments attract the particles each other to form particle chains and increase the transmittance of the suspension, demonstrating a transmissive or bright droplet. In addition, the reflectance of the droplet is measured at various frequencies with different amplitudes.     

Continuous size-based separation of microparticles in a microchannel with symmetric sharp corner structures

A new microchannel with a series of symmetric sharp corner structures is reported for passive size-dependent particle separation. Micro particles of different sizes can be completely separated based on the combination of the inertial lift force and the centrifugal force induced by the sharp corner structures in the microchannel. At appropriate flow rate and Reynolds number, the centrifugal force effect on large particles, induced by the sharp corner structures, is stronger than that on small particles; hence after passing a series of symmetric sharp corner structures, large particles are focused to the center of the microchannel, while small particles are focused at two particle streams near the two side walls of the microchannel. Particles of different sizes can then be completely separated. Particle separation with this device was demonstrated using 7.32 μm and 15.5 μm micro particles. Experiments show that in comparison with the prior multi-orifice flow fractionation microchannel and multistage-multiorifice flow fractionation microchannel, this device can completely separate two-size particles with narrower particle stream band and larger separation distance between particle streams. In addition, it requires no sheath flow and complex multi-stage separation structures, avoiding the dilution of analyte sample and complex operations. The device has potentials to be used for continuous, complete particle separation in a variety of lab-on-a-chip and biomedical applications.     

Continuous sheath-free magnetic separation of particles in a U-shaped microchannel

Particle separation is important to many chemical and biomedical applications. Magnetic field-induced particle separation is simple, cheap, and free of fluid heating issues that accompany electric, acoustic, and optical methods. We develop herein a novel microfluidic approach to continuous sheath-free magnetic separation of particles. This approach exploits the negative or positive magnetophoretic deflection to focus and separate particles in the two branches of a U-shaped microchannel, respectively. It is applicable to both magnetic and diamagnetic particle separations, and is demonstrated through the sorting of 5 μm and 15 μm polystyrene particles suspended in a dilute ferrofluid.