Piezoelectric-driven droplet impact printing with an interchangeable microfluidic cartridge

Microfluidic impact printing has been recently introduced, utilizing its nature of simple device architecture, low cost, non-contamination, and scalable multiplexability and high throughput. In this paper, we have introduced an impact-based droplet printing platform utilizing a simple plug-and-play microfluidic cartridge driven by piezoelectric actuators. Such a customizable printing system allows for ultrafine control of droplet volume from picoliters (∼23 pl) to nanoliters (∼10 nl), a 500 fold variation. The high flexibility of droplet generation can be simply achieved by controlling the magnitude of actuation (e.g., driving voltage) and the waveform shape of actuation pulses, in addition to nozzle size restrictions. Detailed printing characterizations on these parameters have been conducted consecutively. A multiplexed impact printing system has been prototyped and demonstrated to provide the functions of single-droplet jetting and droplet multiplexing as well as concentration gradient generation. Moreover, a generic biological assay has also been tested and validated on this printing platform. Therefore, the microfluidic droplet printing system could be of potential value to establish multiplexed micro reactors for high-throughput life science applications.     

Influences of electric field on living cells in a charged water-in-oil droplet under electrophoretic actuation

We experimentally investigate the effects of high electric field on living cells inside a charged droplet under electrophoretic actuation. When an aqueous droplet suspended in a dielectric liquid contacts with electrified electrode, the droplet acquires charge. This charged droplet undergoes electrophoretic motion under strong electric field (1–3 kV/cm), which can be used as a droplet manipulation method in biomicrofluidic applications. However, because strong electric field and use of dielectric oil can be a harmful environment for living cells, the biological feasibilities have been tested. Trypan blue test and cell growth test have been performed to check the viability and proliferation of cells in a droplet under various electric field strengths and actuation times. We have not observed any noticeable influence of electric field and silicone oil on the viability and proliferation of cells, which indicates that electrophoresis could be safely used as a manipulation method for a droplet containing living biological system.     

Electrowetting on dielectric driven droplet resonance and mixing enhancement in parallel-plate configuration

This study experimentally verifies that the mixing process in a droplet can be enhanced by driving the droplet at resonant frequencies and at alternating driving frequencies using a parallel-plate electrowetting on dielectric device. The mixing time, which is defined as the time required for reaching the well-mixed state, in a resonant droplet is found to be significantly shorter than that in a non-resonant droplet. Besides, it is also found that a higher driving potential leads to a better mixing effect, especially at resonant frequencies. Furthermore, when a droplet is driven by alternating two driving frequencies, especially two resonant frequencies, the mixing efficiency is found to be significantly enhanced for a specific alternating duration of these two frequencies.     

Applications of electrowetting-on-dielectric (EWOD) technology for droplet digital PCR

Digital microfluidics is an elegant technique based on single droplets for the design, composition, and manipulation of microfluidic systems. In digital microfluidics, especially in the electrowetting on dielectric (EWOD) system, each droplet acts as an independent reactor, which enables a wide range of multiple parallel biological and chemical reactions at the microscale. EWOD digital microfluidics reduces reagent and energy consumption, accelerates analysis, enables point-of-care diagnostic, simplifies integration with sensors, etc. Such a digital microfluidic system is especially relevant for droplet digital PCR (ddPCR), thanks to its nanoliter droplets and well-controlled volume distribution. At low DNA concentration, these small volumes allow less than one DNA strand per droplet on average (limited dilution) so that after a fixed number of PCR cycles (endpoint PCR), only the DNA in droplets containing the sequence of interest has been amplified and can be detected by fluorescence to yield an accurate count of the sequences of interest using statistical models. Focusing on ddPCR, this article summarizes the latest development and research on EWOD technology for droplet PCR over the last decade.     

A microfluidic platform for on-demand formation and merging of microdroplets using electric control

We discuss a microfluidic system in which (programmable) local electric fields originating from embedded and protected electrodes are used to control the formation and merging of droplets in a microchannel. The creation of droplets-on-demand (DOD) is implemented using the principle of electrowetting. Combined with hydrodynamic control, the droplet size and formation frequency can be varied independently. Using two synchronized DOD injectors, merging-on-demand (MOD) is achieved via electrocoalescence. The efficiency of MOD is 98% based on hundreds of observations. These two functionalities can be activated independently.     

The dynamics and stability of lubricating oil films during droplet transport by electrowetting in microfluidic devices

The operation of digital microfluidic devices with water droplets manipulated by electrowetting is critically dependent on the static and dynamic stability and lubrication properties of the oil films that separate the droplets from the solid surfaces. The factors determining the stability of the films and preventing surface fouling in such systems are not yet thoroughly understood and were experimentally investigated in this study. The experiments were performed using a standard digital microfluidic cartridge in which water droplets enclosed in a thin, oil-filled gap were transported over an array of electrodes. Stable, continuous oil films separated the droplets from the surfaces when the droplets were stationary. During droplet transport, capillary waves formed in the films on the electrode surfaces as the oil menisci receded. The waves evolved into dome-shaped oil lenses. Droplet deformation and oil displacement caused the films at the surface opposite the electrode array to transform into dimples of oil trapped over the centers of the droplets. Lower actuation voltages were associated with slower film thinning and formation of fewer, but larger, oil lenses. Lower ac frequencies induced oscillations in the droplets that caused the films to rupture. Films were also destabilized by addition of surfactants to the oil or droplet phases. Such a comprehensive understanding of the oil film behavior will enable more robust electrowetting-actuated lab-on-a-chip devices through prevention of loss of species from droplets and contamination of surfaces at points where films may break.     

A model for the determination of the dimensions of dents for jagged electrodes in electrowetting on dielectric microsystems

The motion of a droplet from one electrode to the next in electrowetting on dielectric microsystems is not straightforward. Microfabrication imposes a gap separating the electrodes. This hydrophobic gap has the effect of pinning the triple contact line, preventing the motion of the microdrop. To avoid such a drawback, jagged electrodes, i.e., electrodes with crenellated side boundaries have been designed. In this work, an analytical model for dimensioning the size of the dents of the electrodes is derived based on the contact line elasticity theory. This model determines a lower limit for the nondimensional ratio of the length of a dent to its width. The designs of jagged electrodes in the literature have been verified to satisfy the condition, but with very different safety margins.     

Electrowetting on dielectric device with crescent electrodes for reliable and low-voltage droplet manipulation

Digital microfluidics based on electrowetting on dielectric is an emerging popular technology that manipulates single droplets at the microliter or even the nanoliter level. It has the unique advantages of rapid response, low reagent consumption, and high integration and is mainly applied in the field of biochemical analysis. However, currently, this technology still has a few problems, such as high control voltage, low droplet velocity, and continuity in flow, limiting its application. In this paper, through theoretical analysis and numerical simulation, it is deduced that a drive electrode with a crescent configuration can reduce the driving voltage. The experimental results not only validate this deduction but also indicate that crescent electrode can improve the droplet motion continuity and the success in split rate.     

An electrohydrodynamic flow in ac electrowetting

In ac electrowetting, hydrodynamic flows occur within a droplet. Two distinct flow patterns were observed, depending on the frequency of the applied electrical signal. The flow at low-frequency range was explained in terms of shape oscillation and a steady streaming process in conjunction with contact line oscillation. The origin of the flow at high-frequency range has not yet been explained. We suggest that the high-frequency flow originated mainly from the electrothermal effect, in which electrical charge is generated due to the gradient of electrical conductivity and permittivity, which is induced by the Joule heating of fluid medium. To support our argument, we analyzed the flow field numerically while considering the electrical body force generated by the electrothermal effect. We visualized the flow pattern and measured the flow velocity inside the droplet. The numerical results show qualitative agreement with experimental results with respect to electric field and frequency dependence of flow velocity. The effects of induced-charge electro-osmosis, natural convection, and the Marangoni flow are discussed.     

Coplanar electrowetting-induced stirring as a tool to manipulate biological samples in lubricated digital microfluidics. Impact of ambient phase on drop internal flow pattern

Oscillating electrowetting on dielectrics (EWOD) with coplanar electrodes is investigated in this paper as a way to provide efficient stirring within a drop with biological content. A supporting model inspired from Ko et al. [Appl. Phys. Lett. 94, 194102 (2009)] is proposed allowing to interpret oscillating EWOD-induced drop internal flow as the result of a current streaming along the drop surface deformed by capillary waves. Current streaming behaves essentially as a surface flow generator and the momentum it sustains within the (viscous) drop is even more significant as the surface to volume ratio is small. With the circular electrode pair considered in this paper, oscillating EWOD sustains toroidal vortical flows when the experiments are conducted with aqueous drops in air as ambient phase. But when oil is used as ambient phase, it is demonstrated that the presence of an electrode gap is responsible for a change in drop shape: a pinch-off at the electrode gap yields a peanut-shaped drop and a symmetry break-up of the EWOD-induced flow pattern. Viscosity of oil is also responsible for promoting an efficient damping of the capillary waves which populate the surface of the actuated drop. As a result, the capillary network switches from one standing wave to two superimposed traveling waves of different mechanical energy, provided that actuation frequency is large enough, for instance, as large as the one commonly used in electrowetting applications (f ∼ 500 Hz and beyond). Special emphasis is put on stirring of biological samples. As a typical application, it is demonstrated how beads or cell clusters can be focused under flow either at mid-height of the drop or near the wetting plane, depending on how the nature of the capillary waves is (standing or traveling), and therefore, depending on the actuation frequency (150 Hz–1 KHz).     

Concurrent droplet charging and sorting by electrostatic actuation

This paper presents a droplet-based microfluidic device for concurrent droplet charging and sorting by electrostatic actuation. Water-in-oil droplets can be charged on generation by synchronized electrostatic actuation. Then, simultaneously, the precharged droplets can be electrostatically steered into any designated laminar streamline, thus they can be sorted into one of multiple sorting channels one by one in a controlled fashion. In this paper, we studied the size dependence of the water droplets under various relative flow rates of water and oil. We demonstrated the concurrent charging and sorting of up to 600 droplets∕s by synchronized electrostatic actuation. Finally, we investigated optimized voltages for stable droplet charging and sorting. This is an essential enabling technology for fast, robust, and multiplexed sorting of microdroplets, and for the droplet-based microfluidic systems.     

Fast and reliable droplet transport on single-plate electrowetting on dielectrics using nonfloating switching method

In a droplet transport based on electrowetting on dielectrics, the parallel-plate configuration is more popular than the single-plate one because the droplet transport becomes increasingly difficult without cover plate. In spite of the improved transport performance, the parallel-plate configuration often limits the access to the peripheral components, requesting the removal of the cover plate, the single-plate configuration. We investigated the fundamental features of droplet transport for the single-plate configuration. We compared the performance of several switching methods with respect to maximum speed of successive transport without failure and suggested nonfloating switching method which is inherently free from the charge-residue problem and exerts greater force on a droplet than conventional switching methods. A simple theory is provided to understand the different results for the switching methods.     

Repulsion-based model for contact angle saturation in electrowetting

We introduce a new model for contact angle saturation phenomenon in electrowetting on dielectric systems. This new model attributes contact angle saturation to repulsion between trapped charges on the cap and base surfaces of the droplet in the vicinity of the three-phase contact line, which prevents these surfaces from converging during contact angle reduction. This repulsion-based saturation is similar to repulsion between charges accumulated on the surfaces of conducting droplets which causes the well known Coulombic fission and Taylor cone formation phenomena. In our model, both the droplet and dielectric coating were treated as lossy dielectric media (i.e., having finite electrical conductivities and permittivities) contrary to the more common assumption of a perfectly conducting droplet and perfectly insulating dielectric. We used theoretical analysis and numerical simulations to find actual charge distribution on droplet surface, calculate repulsion energy, and minimize energy of the total system as a function of droplet contact angle. Resulting saturation curves were in good agreement with previously reported experimental results. We used this proposed model to predict effect of changing liquid properties, such as electrical conductivity, and system parameters, such as thickness of the dielectric layer, on the saturation angle, which also matched experimental results.     

A novel actuation method of transporting droplets by using electrical charging of droplet in a dielectric fluid

We evaluate the feasibility of manipulating droplets in two dimensions by exploiting Coulombic forces acting on conductive droplets immersed in a dielectric fluid. When a droplet suspended in an immiscible fluid is located near an electrode under a dc voltage, the droplet can be charged by direct contact, by charge transfer along an electrically conducting path, or by both mechanisms. This process is called electrical charging of droplet (ECOD). This charged droplet may then be transported rapidly by exploiting Coulombic forces. We experimentally demonstrate electrical actuation of a charged droplet by applying voltage sequences. A charged droplet is two dimensionally actuated by following the direction of the electrical field signal. The droplet does not contact the surface of the microfluidic chip when it moves. This characteristic is very advantageous because treatments of the substrate surfaces of microfluidic chip become simpler. In order to test the feasibility of using ECOD in a droplet-based microreactor, electrocoalescence of two oppositely charged droplets is also studied. When two droplets approach each other due to Coulombic attraction, a liquid bridge is formed between them. We postulate that if the applied electric field is weaker than a certain critical level, the two droplets coalesce instantaneously when the charges are exchanged and redistributed through this liquid bridge.     

Novel on-demand droplet generation for selective fluid sample extraction

A novel microfluidic device enabling selective generation of droplets and encapsulation of targets is presented. Unlike conventional methods, the presented mechanism generates droplets with unique selectivity by utilizing a K-junction design. The K-junction is a modified version of the classic T-junction with an added leg that serves as the exit channel for waste. The dispersed phase fluid enters from one diagonal of the K and exits the other diagonal while the continuous phase travels in the straight leg of the K. The intersection forms an interface that allows the dispersed phase to be controllably injected through actuation of an elastomer membrane located above the inlet channel near the interface. We have characterized two critical components in controlling the droplet size-membrane actuation pressure and timing as well as identified the region of fluid in which the droplet will be formed. This scheme will have applications in fluid sampling processes and selective encapsulation of materials. Selective encapsulation of a single cell from the dispersed phase fluid is demonstrated as an example of functionality of this design.     

Versatile on-demand droplet generation for controlled encapsulation

We present a droplet-based microfluidic system for performing bioassays requiring controlled analyte encapsulation by employing highly flexible on-demand droplet generation. On-demand droplet generation and encapsulation are achieved pneumatically using a microdispensing pump connected to a constant pressure source. The system generates single droplets to the collection route only when the pump is actuated with a designated pressure level and produces two-phase parallel flow to the waste route during the stand-by state. We analyzed the effect of actuation pressure on the stability and size of droplets and optimized conditions for generation of stable droplets over a wide pressure range. By increasing the duration of pump actuation, we could either trigger a short train of identical size droplets or generate a single larger droplet. We also investigated the methodology to control droplet contents by fine-tuning flow rates or implementing a resistance bridge between the pump and main channels. We demonstrated the integrated chip for on-demand mixing between two aqueous phases in droplets and on-demand encapsulation of Escherichia coli cells. Our unique on-demand feature for selective encapsulation is particularly appropriate for bioassays with extremely dilute samples, such as pathogens in a clinical sample, since it can significantly reduce the number of empty droplets that impede droplet collection and subsequent data analysis.     

Handling of artificial membranes using electrowetting-actuated droplets on a microfluidic device combined with integrated pA-measurements

Artificial membranes, as a controllable environment, are an essential tool to study membrane proteins. Electrophysiology provides information about the ion transport mechanism across a membrane at the single-protein level. Unfortunately, high-throughput studies and screening are not accessible to electrophysiology because it is a set of not automated and technically delicate methods. Therefore, it is necessary to automate and parallelize electrophysiology measurement in artificial membranes. Here, we present a first step toward this goal: the fabrication and characterization of a microfluidic device integrating electrophysiology measurements and the handling of an artificial membrane which includes its formation, its displacement and the separation of its leaflets using electrowetting actuation of sub-μL droplets. To validate this device, we recorded the insertion of a model porin, α-hemolysin.     

中国低次烟叶资源综合利用研究

中国低次烟叶资源的综合利用不仅关系到烟农的经济效益,同时也关系到资源短缺和环境污染问题的解决.当前中国低次烟叶资源非常丰富,种植面积133×104hm2左右,年产量大约为240×104t,均居世界首位;但正面临着综合利用水平低、途径少、资源浪费非常严重等一系列问题.本文综述了中国低次烟叶资源分布特点、变化趋势及综合利用现状,探讨了低次烟叶资源综合利用的途径:部分低次烟叶重新用于卷烟生产;低次烟叶中蛋白质用于生产普通蛋白食品、功能性食品、香精香料、医药学辅助材料和植物保护剂等;茄尼醇用于合成辅酶Q10、VK、抗癌增效剂SDB等;烟碱用于开发高效、低毒、低残留的无公害农药;氨基酸、有机酸和糖等成分可作为重要的化工原料或用于生产香精香料等;论述了低次烟叶资源综合利用的重大意义,旨在为我国低次烟叶资源的综合利用实践提供理论依据和方法借鉴.     

SAW-driven droplet jetting technology in microfluidic: A review

The introduction of surface acoustic wave (SAW) technology on microfluidics has shown its powerfully controlling and actuating fluid and particle capability in a micro-nano scale, such as fluid mixing, fluid translation, microfluidic pumping, microfluidic rotational motor, microfluidic atomization, particle or cell concentration, droplet or cell sorting, reorientation of nano-objects, focusing and separation of particles, and droplet jetting. The SAW-driven droplet jetting technology enjoys the advantages of simple structure to fabricate with little hindrance, compact size to integrate with other components, high biocompatibility with biological cells or other molecule samples, large force in realizing fast fluidic actuation, and contact-free manipulation with fluid. The realization of this technology can effectively overcome some bottleneck problems in the current micro-injection technology, such as mechanical swear, complicated and bulky structure, and strict limitation of requirements on fluidic characteristics. This article reviews and reorganizes SAW-microfluidic jetting technology from decades of years, referring to the interaction mechanism theory of SAW and fluid, experimental methods of SAW-microfluidic jetting, effects of related parameters on objected pinch-off droplets, and applications of individual structures. Finally, we made a summary of the research results of the current literature and look forward and appraise where this discipline of SAW-microfluidic jetting could go in the future.     

On-chip actuation transmitter for enhancing the dynamic response of cell manipulation using a macro-scale pump

An on-chip actuation transmitter for achieving fast and accurate cell manipulation is proposed. Instead of manipulating cell position by a directly connected macro-scale pump, polydimethylsiloxane deformation is used as a medium to transmit the actuation generated from the pump to control the cell position. This actuation transmitter has three main advantages. First, the dynamic response of cell manipulation is faster than the conventional method with direct flow control based on both the theoretical modeling and experimental results. The cell can be manipulated in a simple harmonic motion up to 130 Hz by the proposed actuation transmitter as opposed to 90 Hz by direct flow control. Second, there is no need to fill the syringe pump with the sample solution because the actuation transmitter physically separates the fluids between the pump and the cell flow, and consequently, only a very small quantity of the sample is required (<1 μl). In addition, such fluid separation makes it easy to keep the experiment platform sterilized because there is no direct fluid exchange between the sample and fluid inside the pump. Third, the fabrication process is simple because of the single-layer design, making it convenient to implement the actuation transmitter in different microfluidic applications. The proposed actuation transmitter is implemented in a lab-on-a-chip system for red blood cell (RBC) evaluation, where the extensibility of red blood cells is evaluated by manipulating the cells through a constriction channel at a constant velocity. The application shows a successful example of implementing the proposed transmitter.