High-throughput sorting of nanoliter droplets enabled by a sequentially addressable dielectrophoretic array

Droplet microfluidics has emerged as a powerful tool for a diverse range of biomedical and industrial applications such as single-cell analysis, directed evolution, and metabolic engineering. In these applications, droplet sorting has been effective for isolating small droplets encapsulating molecules, cells, or crystals of interest. Recently, there is an increased interest in extending the applicability of droplet sorting to larger droplets to utilize their size advantage. However, sorting throughputs of large droplets have been limited, hampering their wide adoption. Here, we report our demonstration of high-throughput fluorescence-activated droplet sorting of 1 nL droplets using an upgraded version of the sequentially addressable dielectrophoretic array (SADA), which we reported previously. The SADA is an array of electrodes that are individually and sequentially activated/deactivated according to the speed and position of a droplet passing nearby the array. We upgraded the SADA by increasing the number of driving electrodes constituting the SADA and incorporating a slanted microchannel. By using a ten-electrode SADA with the slanted microchannel, we achieved fluorescence-activated droplet sorting of 1 nL droplets at a record high throughput of 1752 droplets/s, twice as high as the previously reported maximum sorting throughput of 1 nL droplets.     

Electrical actuation of dielectric droplets by negative liquid dielectrophoresis

This paper presents an electrical actuation scheme of dielectric droplet by negative liquid dielectrophoresis. A general model of lumped parameter electromechanics for evaluating the electromechanical force acting on the droplets is established. The model reveals the influence of actuation voltage, device geometry, and dielectric parameter on the actuation force for both conductive and dielectric medium. Using this model, we compare the actuation forces for four liquid combinations in the parallel-plate geometry and predict the low voltage actuation of dielectric droplets by negative dielectrophoresis. Parallel experimental results demonstrate such electric actuation of dielectric droplets, including droplet transport, splitting, merging, and dispending. All these dielectric droplet manipulations are achieved at voltages < 100 V_rms. The frequency dependence of droplet actuation velocity in aqueous solution is discussed and the existence of surfactant molecules is believed to play an important role by realigning with the AC electric field. Finally, we present coplanar manipulation of oil and water droplets and formation of oil-in-water emulsion droplet by applying the same low voltage.     

Mechanistic studies of droplet electrophoresis: A review

Electrophoresis (EP) of droplets is an intriguing phenomenon that has applications in biological systems, separation strategies, and reactor engineering. Droplet EP is significantly different from the classic particle EP because of droplet characteristics such as a mobile surface charge and the nonrigidity of the interface. Also, the liquid–liquid system, where there is an interplay between the hydrodynamic and electrokinetic forces in both phases, adds to the complexity of electrophoretic motion. Due to the vast amount of potential applications of droplet EP, a mechanistic understanding of the droplet motion in the presence of an external electric field is crucial. This review provides a background on the mechanism of droplet EP and summarizes the intrinsic interplay between the different relevant forces in these systems. The review also describes the key differences between droplet EP and particle EP, and the impact of these differences on droplet mobility. Additionally, we schematically summarize the effects of key parameters on droplet EP mobility, such as electric double layer polarization, the development of internal flow inside a droplet and boundary effects.     

Airborne virus transmission via respiratory droplets: Effects of droplet evaporation and sedimentation

Airborne transmission is considered as an important route for the spread of infectious diseases, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and is primarily determined by the droplet sedimentation time, that is, the time droplets spend in air before reaching the ground. Evaporation increases the sedimentation time by reducing the droplet mass. In fact, small droplets can, depending on their solute content, almost completely evaporate during their descent to the ground and remain airborne as so-called droplet nuclei for a long time. Considering that viruses possibly remain infectious in aerosols for hours, droplet nuclei formation can substantially increase the infectious viral air load. Accordingly, the physical-chemical factors that control droplet evaporation and sedimentation times and play important roles in determining the infection risk from airborne respiratory droplets are reviewed in this article.     

微流控芯片液滴生成与检测技术研究进展

微流控芯片液滴技术是一种操控微小体积液体的新技术,既可实现高通量微观样本的生成及控制,也可进行独立液滴的操作.分散的微液滴单元可作为理想的微反应器,在生物医药中的药物筛选、材料筛选和高附加值微颗粒材料合成领域展现出巨大的应用潜力.液滴微流控芯片是利用流体剪切力的改变,使互不相溶的两相流体在其界面处生成稳定、有序的液滴,...     

The head-on colliding process of binary liquid droplets at low velocity: high-speed photography experiments and modeling

The experimental and theoretical studies are reported in this paper for the head-on collisions of a liquid droplet with another of the same fluid resting on a solid substrate. The droplet on the hydrophobic polydimethylsiloxane (PDMS) substrate remains in a shape of an approximately spherical segment and is isometric to an incoming droplet. The colliding process of the binary droplets was recorded with high-speed photography. Head-on collisions saw four different types of response in our experiments: complete rebound, coalescence, partial rebound with conglutination, and coalescence accompanied by conglutination. For a complete rebound, both droplets exhibited remarkable elasticity and the contact time of the two colliding droplets was found to be in the range of 10–20 ms. With both droplets approximately considered as elastic bodies, Hertz contact theory was introduced to estimate the contact time for the complete rebound case. The estimated result was found to be on the same order of magnitude as the experimental data, which indicates that the present model is reasonable.     

Spontaneous generation of emulsion droplets by autonomous fluid-pumping using the gas permeability of poly(dimethylsiloxane) (PDMS)

Conventional droplet-based microfluidic systems require expensive, bulky external apparatuses, such as electric power supplies and pressure-driven pumps for fluid transportation. This study demonstrates an alternative way to produce emulsion droplets by autonomous fluid-handling based on the gas permeability of poly(dimethylsiloxane) (PDMS). Furthermore, basic concepts of fluid-handling are expanded to control the direction of the microfluid in the microfluidic device. The alternative pumping energy resulting from the high gas permeability of PDMS is used to generate water-in-oil (W/O) emulsions, which require no additional structures apart from microchannels. We can produce emulsion droplets by simple loading of the oil and aqueous solutions into the inlet reservoirs. During the operation of the microfluidic device, changes in droplet size, volumetric flow rate, and droplet generation frequency were quantitatively analyzed. As a result, we found that changes in the wetting properties of the microchannel greatly influence the volumetric flow rate and droplet generation frequency. This alternative microfluidic approach for preparing emulsion droplets in a simple and efficient manner is designed to improve the availability of emulsion droplets for point of care bioanalytical applications, in situ synthesis of materials, and on-site sample preparation tools.     

水滴在超疏水植物叶片上的沉积方法和机理研究进展

自然界中有很多超疏水植物叶片, 水滴撞击在这些表面时极易产生溅射和反弹, 造成农用化学品喷雾施药时药物的大量损失, 利用率低下, 从而重复喷洒施药. 农用化学品过度使用将造成食品安全、 农药残留、 水资源浪费及环境生态污染等问题. 因此, 增加水滴在超疏水植物叶片表面的沉积效率对提高农药利用率尤为重要. 本文从分析水滴在超疏水表面的撞击动力学特征开始, 结合添加助剂后液滴的物理化学性质, 系统阐述了水滴在超疏水植物叶片上的沉积方法和机理, 并提出筛选助剂和研究机理不仅要考虑助剂性质还要结合基底结构、 撞击动力学特征等因素, 而且还要考虑单水滴尺寸大小、 基底运动和弹性及环境因素等对沉积的影响. 本文对农药喷洒及生物医学、 机械工程、 涂料喷涂和油墨打印等领域均有指导意义和应用价值.     

A homogeneous assay for protein analysis in droplets by fluorescence polarization

We present a novel homogeneous (“mix‐incubate‐read”) droplet microfluidic assay for specific protein detection in picoliter volumes by fluorescence polarization (FP), for the first time demonstrating the use of FP in a droplet microfluidic assay. Using an FP‐based assay we detect streptavidin concentrations as low as 500 nM and demonstrate that an FP assay allows us to distinguish droplets containing 5 μM rabbit IgG from droplets without IgG with an accuracy of 95%, levels relevant for hybridoma screening. This adds to the repertoire of droplet assay techniques a direct protein detection method which can be performed entirely inside droplets without the need for labeling of the analyte molecules.     

流动聚焦型微流控体系中的液滴的图案化排列和转变模式

流体在微流通道中形成剪切流场（低雷诺数）．不同于宏观体系,由于剪切力和表面张力的竞争作用,产生的液滴在微尺度下的微流通道中形成特殊的排列现象---周期性类似“晶格”排列现象．设计了新型流动聚焦型微流控芯片,分析研究在微流体系中液滴周期性图案化排列和转变机理性,液滴排列模式受两方面因素影响：水油两相的流速比值和微通道尺寸．当微通道宽度为250或300 μm时,液滴形成单层分散,双层和单层挤压排列．当微通道宽度为350 μm 时,液滴会形成单层分散到三层排列到双层挤压最后到单层挤压排列．当出口通道宽度增加到400 μm时,甚至出现了液滴四层排列的现象．同时研究了各个液滴排列模式的“转变点”.     

Partitioning invertase between a dilute water solution and generated droplets

Water droplets or mist occur naturally in the air at seashores. These water droplets carry inorganic and organic substances from the sea to the land via the air, creating fertile land in sandy coastal areas (1). The same phenomenon occurs in an air-fluidized bed bioreactor (2). In an air-fluidized bed reactor, proteins can be transferred from the bioreactor semisolid bulk phase to an enriched droplet phase. This protein transfer process (droplet fractionation) can be experimentally simulated by shaking a separatory funnel containing a dilute solution of a given protein, which can be an enzyme like invertase. The created droplets become richer in invertase (protein) than that of the original dilute solution. The droplets can then be coalesced by tranpping them and recovering the concentrated protein in the new liquid phase. Typically, in such a droplet fractionation process a collected enzyme can be degraded in its ability tocatalyze a chemical reaction. In this article, we explore whether the initial solution pH control variable can be adjusted to minimize the decrease of enzyme activity in this process. The protein droplet recovery problem is one in which the recovered amount of desired protein (enzyme) in the droplet is maximized, subject to the minimization of the enzyme activity loss. The partition coefficient, which is the ratio between the protein concentration in the droplets and the residual solution, is maximized at approx 4.8 and occurs at pH 3.0. Here, the partition coefficient for invertase decreases as the initial solution pH increases, between pH 3.0 and 8.0. Interestingly, the initial solution surface tension seems to be inversely proportional to the partition coefficient. The partition coefficien treachesa maximum value at a surface tension value of approx 63 mN/m at pH 3.0. The enzymatic activity of the initial, the residual, and the droplet solutions all decrease as the bulksolution pH increases. Adecrease of enzymatic activity was observed in the residual bulk solution when compared with that in the initial bulk solution at all pH levels. Also, up to 90% of the invertase activity was lost in the droplets when compared to the initial bulk solution.     

Electrochemical Detection of Droplets in Microfluidic Devices: Simultaneous Determination of Velocity,Size and Content

Microfluidic devices were designed to electrochemically detect in a two‐phase flow the velocity, size and content of aqueous droplets containing redox species. The principle of these determinations is based on the analysis of a unique chronoamperometric response recorded during the passage of a droplet over channel microelectrodes. Two configurations of electrochemical cell with different geometries were investigated both theoretically and experimentally. Velocity and size of droplets, as well as internal recirculating convection within droplets, were evaluated from chronoamperometric curves by specific transition times depending on the cell configuration. In addition, the droplet content was probed from the Faradaic current controlled by mass transport and by internal hydrodynamic regime. For droplet velocity and size, experimental data were systematically compared to optical measurements. All the results demonstrated the high performance of the electrochemical detection reached under these conditions. They successfully validate the concept of self‐consistent electrochemical detections of aqueous droplets within microchannels for the simultaneous determination of their velocity, size and content.     

Formation of Concentrated Nanoemulsions by Extreme Shear

Abstract

We have systematically investigated the production of “nanoemulsions,” droplets of one liquid phase in another immiscible liquid phase that have diameters less than 100 nm. Our approach relies on a combination of extreme shear due to multipass, high‐pressure microfluidic injection and systematic control of the emulsion's composition. By repeatedly shearing a silicone oil‐in‐water emulsion in an inhomogeneous extensional shear flow, the multipass approach enables us to reduce the droplet polydispersity and average radius. Using dynamic light scattering, we study the changes in the average radius, ?a?, as a function of the number of passes, driving injection pressure (i.e., shear rate), droplet volume fraction, surfactant concentration, and droplet oil viscosity. The smallest nanoemulsion that we obtain has ?a?=18 nm. At large droplet volume fractions φ≥0.65, we observe phase inversion, rather than a reduction in the droplet size. This provides evidence that droplet coalescence can occur during extreme shear, even when a significant excess of a strongly stabilizing surfactant is present.     

Controlled generation of droplets using an electric field in a flow-focusing paper-based device

Droplet-based microfluidics is a modular platform in high-throughput single-cell and small sample analyses. However, this droplet microfluidic system was widely fabricated using soft lithography or glass capillaries, which is expensive and technically demanding for various applications, limiting use in resource-poor settings. Besides, the variation in droplet size is also restricted due to the limitations on the operating forces that the paper-based platform is able to withstand. Herein, we develop a fully integrated paper-based droplet microfluidic platform for conducting droplet generation and cell encapsulation in independent aqueous droplets dispersed in a carrier oil by incorporating electric fields. Through imposing an electric field, the droplet size would decrease with increasing the electric field and smaller droplets can be produced at high applied voltage. The droplet diameter can be adjusted by the ratio of inner and outer flow velocities as well as the applied electric field. We also demonstrated the proof of concept encapsulation application of our paper device by encapsulating yeast cells under an electric field. Using a simple wax printing method, carbon electrodes can be integrated on the paper. The integrated paper-based microfluidic platform can be fabricated easily and conducted outside of centralized laboratories. This microfluidic system shows great potential in drug and cell investigations by encapsulating cells in resource-limited environments.     

Plateau Rayleigh instability simulation

The well-known phenomena of Plateau-Rayleigh instability has been simulated using computational fluid dynamics (CFD). The breakup of a liquid film into an array of droplets on a cylindrical element was simulated using a volume-of-fluid (VOF) solver and compared to experimental observations and existing theory. It is demonstrated that the VOF method can correctly predict the breakup of thins films into an array of either axisymmetric droplets or clam-shell droplets, depending on the surface energy. The existence of unrealistically large films is precluded. Droplet spacing was found to show reasonable agreement with theory. Droplet motion and displacement under fluid flow was also examined and compared to that in previous studies. It was found that the presence of air flow around the droplet does not influence the stable film thickness; however, it reduces the time required for droplet formation. Novel relationships for droplet displacement were derived from the results.     

液体弹珠：制备策略、物理性质及应用探索

Liquid marbles (LMs) are liquid droplets coated with a layer of lyophobic particles at the air-liquid interface. Since the pioneering work by Aussillous et al. in 2001, LMs have attracted significant attention owing to their facile fabrication, flexibility in the choice of the constituent particles and liquids, intriguing properties such as non-wetting and non-adhesive nature, satisfactory elasticity and stability, as well as promising applications in microfluidics, sensors, controlled release, and microreactors. The classical strategy for the preparation of LMs involves rolling a small volume of a droplet on a lyophobic powder bed for complete encapsulation of the liquid by the particles. In addition, various innovative methods, including electrostatic and coalescent approaches, have been developed for preparing special LMs with a complicated structure or morphology. Diverse materials such as water, surfactant solutions, liquid metals, reagents, blood, and even viscous adhesives have been employed as the internal liquid for the fabrication of LMs. Theoretically, any particulates such as lycopodium, polytetrafluoroethylene, Fe₃O₄, SiO₂, and graphite grains can be employed as the outer coating, but they are usually required to be lyophobic with sizes of less than hundreds of microns. The unique structure of the particle-covered droplet and the dual solid-liquid characteristics endow LMs with some unique and interesting properties, especially the non-wetting and non-adhesive nature. As the lyophobic coating particles restrain the internal liquid from contacting the substrate, LMs can move easily across either solid or liquid surfaces, neither wetting the substrate nor contaminating the internal liquid. An equally fascinating property of LMs is their satisfactory stability, which is necessary for most of their applications. The high stability of LMs stems from the protection of the coating powders and is embodied in both good mechanical stability (remaining intact after being released from a certain height or under a certain compression) and long lifetime (greatly suppressing the evaporation of the internal liquid). These extraordinary properties make LMs promising candidates for use in multitudinous fields, especially droplet microfluidics and microreactors. The potential application of LMs in microfluidics is ascribed to their non-wetting, non-adhesive nature and other features such as an ability to float on a liquid surface, coalescence, split, a small force of rolling friction, and response to external forces. Notably, LMs hold great promise for applications in microreactions, because they can create a confined reaction microenvironment, minimize reagent usage, facilitate unhindered gas exchange between the internal liquid medium and the surrounding environment, and allow the entry/exit of the reactants/products. We herein review the recent advances in LMs, such as manufacturing techniques, formation mechanisms, physical properties, and emerging applications. In particular, much attention is paid to the factors affecting the stability of LMs and the potential strategies to increase their stability. Moreover, this review discusses the challenges in the future development of LMs, suggests several possible ways of addressing these challenges, and forecasts the future development directions. We believe that this review can help researchers gain a better understanding of LMs and promote their further advances.     

Pumpless dispensing of a droplet by breaking up a liquid bridge formed by electric induction

Dispensing uniform pico‐to‐nanoliter droplets has become one of essential components in various application fields from high‐throughput bio‐analysis to printing. In this study, a new method is suggested and demonstrated for dispensing a droplet on the top plate with an inverted geometry by using electric field. The process of dispensing droplets consists of two stages: (i) formation of liquid bridge by moving up the charged fluid mass using the electrostatic force between the charges on the fluid mass and the induced charges on the substrate and (ii) its break‐up by the motion of the top plate. Different from conventional electrohydrodynamic methods, electric induction enables the droplets to be dispensed on various surfaces including non‐conducting substrate. The use of capillarity with an inverted geometry removes the need of external pumps or elaborates control for constant flow feed. The droplet diameter has been characterized as a function of the nozzle‐to‐plate distance and the plate moving velocity. The robustness of the present method is shown in terms of nozzle length and applied voltage. Finally, its practical applicability is confirmed by rendering a 19 by 24 array of highly uniform droplets with only 1.8% size variation without use of any active feedback control.     

Analysis of the relationship between liquid droplet size and contact angle

The purpose of this paper is to present a consistent theoretical concept that can explain the various physical phenomena associated with the effect of droplet size on contact angle for droplets on solid surfaces, and with the geometry of the liquid/gas/solid contact line in general. Two droplet geometries have been considered: uniformly elongated droplets and axisymmetric droplets. It has been shown that the contact angle for elongated droplets is size-independent and, thus, satisfies the Young equation for constant material and interfacial properties. On the other hand, whereas the contact angle for axisymmetric droplets is size-dependent and does not satisfy the original Young equation, it is shown that this contact angle can still be predicted for any combination of droplet and substrate materials, and a given mass of the droplet. The theoretical work has been combined with the development of numerical schemes of solving the Laplace-Young equation for various droplet geometries. The proposed approach has been applied to different material/substrate combinations and validated against several sets of experimental data. As a result, a method has been developed for predicting the contact angle of both long and axisymmetric sessile droplets of arbitrary sizes for given liquid/solid/gas properties.     

Simultaneous generation of multiple aqueous droplets in a microfluidic device

This paper describes a microfluidic platform for the on-demand generation of multiple aqueous droplets, with varying chemical contents or chemical concentrations, for use in droplet based experiments. This generation technique was developed as a complement to existing techniques of continuous-flow (streaming) and discrete-droplet generation by enabling the formation of multiple discrete droplets simultaneously. Here sets of droplets with varying chemical contents can be generated without running the risk of cross-contamination due to the isolated nature of each supply inlet. The use of pressure pulses to generate droplets in parallel is described, and the effect of droplet size is examined in the context of flow rates and surfactant concentrations. To illustrate this technique, an array of different dye-containing droplets was generated, as well as a set of droplets that displayed a concentration gradient of a fluorescent dye.     

A new droplet breakup phenomenon in electrokinetic flow through a microchannel constriction

A completely new droplet breakup phenomenon is reported for droplets passing through a constriction in an electrokinetic flow. The breakup occurs during the droplet shape recovery process past the constriction throat by the interplay of the dielectrophoretic stress release and the interface energy for droplets with smaller permittivity than that of the ambient fluid. There are conditions for constriction ratios and droplet size that the droplet breakup occurs. The numerical predictions provided here require experimental verification, and then can give rise to a novel microfluidic device design with novel droplet manipulations.