Scalable attoliter monodisperse droplet formation using multiphase nano-microfluidics

We demonstrate a robust method to produce monodisperse femtoliter to attoliter droplets by using a nano-microfluidic device. Two immiscible liquids are forced through a nanochannel where a steady nanoscopic liquid filament forms, thinning close to the nanochannel exit to a microchannel due to the capillary focusing. When the nanoscopic filament enters the microchannel, monodisperse droplets are formed by capillary instability. In a certain range of physical parameters and geometrical configurations, the droplet size is only determined by the nanochannel height and independent of liquid flow rates and ratios, surfactants, and continuous phase viscosity. By using nanochannels with a height of 100–900 nm, 0.4–3.5 μm diameter droplets (volume down to 30 aL) have been produced. The generated droplets are stable for at least weeks.     

Effect of operating conditions and liquid physical properties on the size of monodisperse microbubbles produced in a capillary embedded T-junction device

The aim of this study was to investigate the effect of operating parameters such as liquid flow rate, gas inlet pressure, and capillary diameter as well as the influence of the physical properties of the liquid, in particular viscosity, on the generation of monodisperse microbubbles in a circular cross section T-junction device. Aqueous glycerol solutions with viscosities ranging from 1- to 100 mPa s were used in the experiments. The bubble diameter generated was studied for systematically varied combinations of gas inlet pressure, liquid flow rate, and liquid viscosity with a fixed capillary inner diameter of 150 μm for the liquid and gas inlet channels as well as the outlet channel. In addition, the effect of channel geometry on bubble size was studied using capillaries with inner diameters of first 100 and then 200 μm. In all the experiments the distance between the coaxial capillaries at the junction was set to be 200 μm. All the microbubbles produced in this study were highly monodisperse (polydispersity index <1 %) and it was found, as expected, that bubble formation and size were influenced by the ratio of liquid to gas flow rate, capillary size, and liquid viscosity. The experimental data were then compared with empirical scaling laws derived for rectangular cross-section junctions. In contrast with these previous studies, which have found bubble size to be dependent on either the flow rate ratio (the squeezing regime) or capillary number (the dripping regime), in this experimental study bubble size was found to depend on both capillary number and flow ratio.     

Study of flow behaviors of droplet merging and splitting in microchannels using Micro-PIV measurement

Droplet merging and splitting are important droplet manipulations in droplet-based microfluidics. However, the fundamental flow behaviors of droplets were not systematically studied. Hence, we designed two different microstructures to achieve droplet merging and splitting respectively, and quantitatively compared different flow dynamics in different microstructures for droplet merging and splitting via micro-particle image velocimetry (micro-PIV) experiments. Some flow phenomena of droplets different from previous studies were observed during merging and splitting using a high-speed microscope. It was also found the obtained instantaneous velocity vector fields of droplets have significant influence on the droplets merging and splitting. For droplet merging, the probability of droplets coalescence (η) in a microgroove is higher (50% < η < 92%) than that in a T-junction microchannel (15% < η < 50%), and the highest coalescence efficiency (η = 92%) comes at the two-phase flow ratio e of 0.42 in the microgroove. Moreover, compared with a cylinder obstacle, Y-junction bifurcation can split droplets more effectively and the droplet flow during splitting is steadier. The results can provide better understanding of droplet behaviors and are useful for the design and applications of droplet-based microfluidics.     

Numerical simulation of the capillary flow in the meander microchannel

Pumping in microfluidic devices is an important issue in actuating fluid flow in microchannel, especially that capillary force has received more and more attractions due to the self-driven motion without external power input. However, less 2D simulation was done on the capillary flow in microchannel especially the meander microchannel which can be used for mixing and lab-on-a-chip (LOC) application. In this paper, the numerical simulation of the capillary flow in the meander microchannel has been studied using computer fluid dynamic simulation software CFD-ACE⁺. Different combinations of channel width in the X-direction denoted as W_x and Y-direction denoted as W_y were designed for simulating capillary flow behavior and pressure drop. The designed four types of meander microchannels (W_x × W_y) were 100 × 100 μm, 100 × 200 μm, 50 × 200 μm, and 50 × 400 μm. In this simulation results, it is found that the capillary pumping speed is highly depending on the channel width. The large speed change occurs at the turning angle of channel width change from W_x to W_y. The fastest pumping effect is found in the meander channel of 100 × 100 μm, which has an average pumping speed of 0.439 mm/s. The slowest average flow speed of 0.205 mm/s occurs in the meander channel of 50 × 400 μm. Changing the meander channel width may vary the capillary flow behavior including the pumping speed and the flow resistance as well as pressure drop which will be a good reference in designing the meander microchannels for microfluidic and LOC application.     

“From microtiter plates to droplets” tools for micro-fluidic droplet processing

Droplet-based microfluidic allows high throughput experimentation in with low volume droplets. Essential fluidic process steps are on the one hand the proper control of the droplet composition and on the other hand the droplet processing, manipulation and storage. Beside integrated fluidic chips, standard PTFE-tubings and fluid connectors can be used in combination with appropriate pumps to realize almost all necessary fluidic processes. The segmented flow technique usually operates with droplets of about 100–500 nL volume. These droplets are embedded in an immiscible fluid and confined by channel walls. For the integration of segmented flow applications in established research workflows—which are usually base on microtiter plates—robotic interface tools for parallel/serial and serial/parallel transfer operations are necessary. Especially dose–response experiments are well suited for the segmented flow technique. We developed different transfer tools including an automated “gradient take-up tool” for the generation of segment sequences with gradually changing composition of the individual droplets. The general working principles are introduced and the fluidic characterizations are given. As exemplary application for a dose–response experiment the inhibitory effect of antibiotic tetracycline on Escherichia coli bacteria cultivated inside nanoliter droplets was investigated.     

Formation of fully closed microcapsules as microsensors by microfluidic double emulsion

Microcapsules templated from microfluidic double emulsions attract a great attention due to their broad new potential applications. We present a method to form transparent polymer microcapsules in small sizes of ~30 μm with aqueous cores and fully closed shells. We controlled the size ratio of the aqueous core to the polymer shell not only by flow rates of the double emulsions, but also by synergetic interaction between surfactants at the interface of immiscible fluids. We also found that fully closed shells can be formed by generating the double emulsion droplets in a jetting regime, in which the aqueous cores are confined centrally in the double emulsion droplets. We demonstrated the formation of barcodes in these microcapsules for multiplexed bioassays. These transparent microcapsules also have wide and high potentials for the development of various microsensors by functionalizing the liquid-state cores with compounds sensitive and responsive to temperature, light or electromagnetic field.     

Bistability in the hydrodynamic resistance of a drop trapped at a microcavity junction

We investigate the flow resistance of a droplet trapped at a constriction in a microcavity located at a microchannel bifurcation as a function of system parameters including capillary number, drop confinement, and viscosity ratio. Using a combination of experiments and volume-of-fluid numerical simulations, we measure the hydrodynamic resistance of the trapped drop and connect it to drop deformation in the microcavity. For drop sizes smaller than the microcavity, we observe a bistable behavior in terms of the resistance of the trapped drop as a function of capillary number. For these underfilled drops, we find that the resistance is low at small capillary number (Ca < 10^?3) and jumps to high resistance at a threshold capillary number. For drops equal to the microcavity size, we observe that the bistability vanishes and the drop resistance is of similar magnitude as that of underfilled drops at large capillary number. To explain these findings, we use confocal microscopy and simulations to obtain three-dimensional views of the drop deformation and continuous phase fluid in the microcavity. We observe that the low resistance is due to negligible drop deformation and unobstructed flow of continuous phase through the constriction. The high resistance is due to the drop interface protruding into the constriction restricting the flow of continuous phase through the gutters. Taken together, our results indicate that a trapped drop at a bifurcation can act as a nonlinear resistor and could be potentially used as a soft switch to control droplet trajectories in microfluidic devices.     

Contactless sensing of the conductivity of aqueous droplets in segmented flow

We present an electrode arrangement for the inline measurement of the conductivity of droplets in segmented flow by impedance spectroscopy. We use a thin-walled glass capillary with electrodes contacting the outer surface, so that the contactless measurement of conductivity of the liquid within the capillary is possible. The surface of the glass capillary is silanized resulting in a single hydrophobic surface across which droplets can freely move. We model the impedance of such insulated electrodes and use the model to optimize the electrode system. Measurement of solutions with various salt concentrations allows the performance of the electrode structure to be characterized. Subsequently, the measurement of the impedance response of the aqueous segments in two-phase flow was demonstrated. Measurements were firstly performed with an impedance analyzer and subsequently with a multi-sine measurement setup that is better suited to high-speed measurement of droplets. Previous electrical measurements of segmented flow sensed the difference in dielectric constant between the aqueous phase and the carrier fluid through measurement of capacitance. This work describes an electrical measurement of the conductivity of droplets in segmented flow, that is, the sensor senses a variable property of the droplet itself.     

Nebulization of water/glycerol droplets generated by ZnO/Si surface acoustic wave devices

Efficient nebulization of liquid sessile droplets (water and water/glycerol mixtures) was investigated using standing waves generated using ZnO/Si surface acoustic wave (SAW) devices under different RF powers, frequencies and liquid viscosity (varied glycol concentrations in water). At such high RF powers, there are strong competitions between vertical jetting and nebulization. At lower SAW frequencies of 12.3 and 23.37 MHz, significant capillary waves and large satellite droplets were generated before nebulization could be observed. At frequencies between 23.37 and 37.2 MHz, spreading, displacement or occasionally jetting of the parent sessile droplet was frequently observed before a significant nebulization occurred. When the SAW frequencies were increased from 44.44 to 63.3 MHz, the minimum RF power to initiate droplet nebulization was found to increase significantly, and jetting of the parent droplet before nebulization became significant, although the average size of the nebulized particles and ejected satellite droplets appeared to decrease with an increase in frequency. With the increase of glycerol concentration in the test sessile droplets (or increase in liquid viscosity), nebulization became difficult due to the increased SAW damping rate inside the liquid. Acoustic heating effects were characterized to be insignificant and did not show apparent contributions to the nebulization process due to silicon substrate’s natural effect as an effective heat sink and the employment of a metallic holder beneath the ZnO/Si SAW device substrates.     

Generating gas/liquid/liquid three-phase microdispersed systems in double T-junctions microfluidic device

This article describes the generation of microdispersed bubbles and droplets in a double T-junctions microfluidic device to form immiscible gas/liquid/liquid three-phase flowing systems. Segmented gas plugs are controllably prepared in water at the first T-junction to form gas/liquid two-phase fluid with the perpendicular flow cutting method. Then using this two-phase fluid as the cross-shearing fluid for the oil phase at the second T-junction, the gas/liquid/liquid three-phase flowing systems are prepared. Interestingly, it is found that the break-up of the oil droplets is mainly dominated by the cutting effect of the gas/liquid interface or the pressure drop across the emerging droplet, but independent with the viscous shearing effect of the continuous phase, even at the capillary number (Ca = u _wμ_w/γ_ow) higher than 0.01. The size laws and the distributions of the bubbles and droplets are investigated carefully, and a mathematical model has been developed to relating the operating conditions with the dispersed sizes.     

Droplets coalescence and mixing with identical and distinct surface tension on a wettability gradient surface

The influence of identical and distinct surface tensions on the coalescence and mixing of droplets after a direct collision on a wettability gradient surface (made from a self-assembled monolayer, SAM technique) was investigated. The results indicate that their mixing is driven sequentially by interior convection and diffusion; the convection endures less than 100 ms but dominates more than 60 % of the mixing. If the stationary droplet has a large surface tension (73.28 mN × m^?1), whether the moving droplet has a large surface tension (73.28 mN × m^?1) or a small surface tension (38.63 mN × m^?1), the mushroom-shaped mixing pattern is generated within the coalesced droplet that enhances the convective mixing and also significantly enlarges the interface for mass diffusion. The mixing index of these two cases was greater than 0.8 at 120 s after the collision. For the cases in which the stationary droplet with a small surface tension collided by the moving droplet with a large surface tension, a mixing pattern with a round-head shape developed, which was insufficient to benefit the mixing. When the stationary and moving droplets both had small surface tension, the moving droplet was unable to merge with stationary droplet and had poor mixing quality due to the small surface Gibbs energy of both stationary and moving droplets. For the collision of droplets of identical surface tension, the surface tension affects the coalescence behavior; for the collision of droplets with distinct surface tension, the coalescence behavior and mixing quality depend on the colliding arrangement of stationary and moving droplets.     

Using a cross-flow microfluidic chip for monodisperse UV-photopolymerized microparticles

In this paper, we report a microfluidic chip containing a cross-junction channel for the manipulation of UV-photopolymerized microparticles. Hydrodynamic-focusing is used to form a series of using 365 nm UV light to solidify the hydrogel droplets. We were able to control the size of the hydrogel droplets from 75 to 300 μm in diameter by altering the sample and by changing the flow rate ratio of the mineral oil in the center inlet channel to that of the side inlet channels. We found that the size of the emulsions increases with an increase in average velocity of the dispersed phase flow (polymer solution flow). The size of the emulsions decreases with an average velocity increase of the continuous phase flow (mineral oil flow). Experimental data show that the emulsions are very uniform. The developed microfluidic chip has the advantages of ease of fabrication, low cost, and high throughput. The emulsions generated are very uniform and have good regularity.     

Microchannel emulsification for mass production of uniform fine droplets: integration of microchannel arrays on a chip

We present a novel microchannel emulsification (MCE) system for mass-producing uniform fine droplets. A 60 × 60-mm MCE chip made of single-crystal silicon has 14 microchannel (MC) arrays and 1.2 × 10⁴ MCs, and each MC array consists of many parallel MCs and a terrace. A holder with two inlet through-holes and one outlet through-hole was also developed for simply infusing each liquid and collecting emulsion products. The MCE chip was sealed well by physically attaching it to a flat glass plate in the holder during emulsification. Uniform fine droplets of soybean oil with an average diameter of 10 μm were reliably generated from all the MC arrays. The size of the resultant fine droplets was almost independent of the dispersed-phase flow rate below a critical value. The continuous-phase flow rate was unimportant for both the droplet generation and the droplet size. The MCE chip enabled mass-producing uniform fine droplets at 1.5 ml h⁻¹ and 1.9 × 10⁹ h⁻¹, which could be further increased using a dispersed phase of low viscosity.     

On-chip encapsulation via chaotic mixing

A microfluidic device for production of uniform size capsules with a prescribed membrane thickness is described. It is versatile, novel and suitable for various polymerization reactions. Parameters such as polymerization time and reagent concentrations can be precisely tuned to control the membrane properties. The device features a part which allows to overcome the diffusion barrier by initiating interfacial polymerization via chaotic mixing. It also allows the termination of the reaction and the collection of the resulting capsules. We observe different typical dynamical phenomena occurring in capsules during their flow along the microchannel, namely wrinkling of the membrane, parachute and bullet shapes and bursting of the capsules due to strong hydrodynamical flow. In addition to production, the monitoring of capsule dynamics in flow gave a possibility to estimate the elastic surface modulus \(E_{{\rm S}}\) and the membrane thickness t. We found that \(E_{{\rm S}}\) can be as low as 6 × 10^?3 N m^?1 and that the thickness can be below 100 nm. This microfluidic device is therefore capable of producing uniform size capsule solutions with suitable membrane properties for the controlled release of drugs, and as a model system of red blood cells for microhydrodynamics experiments.     

Micro-PIV investigation of the internal flow transitions inside droplets traveling in a rectangular microchannel

This paper discusses the studies on the internal flow field of droplets traveling in a rectangular microchannel by means of microparticle image velocimetry, specifically concentrating on the effects of capillary number, viscosity ratio and interfacial tension. The flow topology is predominantly dependent on the capillary number. It shows that the evident transitions from three pairs of recirculation zones at lower capillary numbers to one pair of recirculation zones near the sidewalls with low velocity in the central area at intermediate capillary numbers, then to a pair of recirculation zones closest to the axial centerline with high velocity in the central area at higher capillary numbers. There are two critical capillary numbers increasing with viscosity ratio in the evolution of flow features. Droplet size only influences two velocity components values other than the flow topology within intervals separated by the critical values. The equilibrium mechanism of viscous friction force and Marangoni stress dominate the internal topological transition in a surfactant added system. The obtained internal fluid phenomena inside droplets are beneficial to provide a guideline for screening of biochemical reaction conditions in the device.     

Arrays of high-aspect ratio microchannels for high-throughput isolation of circulating tumor cells (CTCs)

Microsystem-based technologies are providing new opportunities in the area of in vitro diagnostics due to their ability to provide process automation enabling point-of-care operation. As an example, microsystems used for the isolation and analysis of circulating tumor cells (CTCs) from complex, heterogeneous samples in an automated fashion with improved recoveries and selectivity are providing new opportunities for this important biomarker. Unfortunately, many of the existing microfluidic systems lack the throughput capabilities and/or are too expensive to manufacture to warrant their widespread use in clinical testing scenarios. Here, we describe a disposable, all-polymer, microfluidic system for the high-throughput (HT) isolation of CTCs directly from whole blood inputs. The device employs an array of high aspect ratio (HAR), parallel, sinusoidal microchannels (25 × 150 μm; W × D; AR = 6.0) with walls covalently decorated with anti-EpCAM antibodies to provide affinity-based isolation of CTCs. Channel width, which is similar to an average CTC diameter (10–20 μm), plays a critical role in maximizing the probability of cell/wall interactions and allows for achieving high CTC recovery. The extended channel depth allows for increased throughput at the optimized flow velocity (2 mm/s in a microchannel); maximizes cell recovery, and prevents clogging of the microfluidic channels during blood processing. Fluidic addressing of the microchannel array with a minimal device footprint is provided by large cross-sectional area feed and exit channels poised orthogonal to the network of the sinusoidal capillary channels (so-called Z-geometry). Computational modeling was used to confirm uniform addressing of the channels in the isolation bed. Devices with various numbers of parallel microchannels ranging from 50 to 320 have been successfully constructed. Cyclic olefin copolymer (COC) was chosen as the substrate material due to its superior properties during UV-activation of the HAR microchannels surfaces prior to antibody attachment. Operation of the HT-CTC device has been validated by isolation of CTCs directly from blood secured from patients with metastatic prostate cancer. High CTC sample purities (low number of contaminating white blood cells) allowed for direct lysis and molecular profiling of isolated CTCs.     

Microchannel geometry design for rapid and uniform reagent distribution

Rapid and uniform reagent distribution is critical to the performance of a high-throughput microfluidic system, and its geometric design of the microchannels dominates the accuracy and distribution uniformity of the daughter droplets. This research’s purpose is to optimize the geometry of the T-junction to achieve a uniform distribution of two daughter droplets from a single liquid droplet. Models of gas–liquid flow were realized in the transient numerical simulations to investigate the geometry-dependent pressure distributions and the flowing velocities inside the droplet during the splitting process that leads to an improved design of the T-junction that can increase the stability of the droplet splitting process. To validate that increasing the stability of the splitting process can help improve the distribution uniformity of the daughter droplets, microfluidic devices were manufactured on poly(methyl methacrylate) substrates with micromilling and thermal bonding for experiments. In the multiple experiments, 2 μl of reagent was loaded into the microfluidic device and a uniform pneumatic pressure was applied to push the droplet into the T-junction for splitting. The experimental results, after statistical analysis, show that the improved T-junction can achieve better distribution uniformity of the daughter droplets with a higher reliability and a less reagent loss during the splitting process.     

Characterization of a miniaturized liquid bridge for nL sample infusion: a comparative study of sample flush-out behavior using flow simulations and direct ESI-MS analysis

In this work we shed light on the microfluidics of a miniaturized liquid bridge that forms the central part of a so-called “capillary gap sampler,” a novel device for rapid and seamless injection of nanoliter sample volumes into an electrospray ionization mass spectrometer (ESI-MS). Parameters relevant for sample flush-out at the liquid bridge and in the spray capillary were identified by systematic variation of the capillary dimensions, the linear buffer flow rate (2.1–34 mm/s) and molecular weight of the analytes (0.5–30 kDa). We found that a reduction in capillary wall thickness by a factor of 1.6 significantly influences analyte peak shapes, leads to an inversion of the relationship between peak width and analyte molecular weight, and allows a fivefold decrease in peak width for large molecules down to 5 s. The results could be verified and explained by simulations, in which the presence of diffusion-controlled “dead zones” at the liquid bridge and dispersion in the spray tip that depend on analyte molecular weight were identified as key factors relevant for the sample flush-out process. The merging of simulations and experimental data gives useful hints toward the re-design of a spray tip as built-in ESI-MS interface for an optimized gap sampler performance.     

Fast fabrication of microfluidic devices using a low-cost prototyping method

Conventional ways to produce microfluidic devices cost a lot due to the requirements for cleanroom environments and expensive equipment, which prevents the wider applications of microfluidics in academia and in industry. In this paper, a dry film photoresist was utilized in a simple way to reduce the fabrication cost of microfluidic masters. Thus, a fast prototyping and fabrication of microstructures in polydimethylsiloxane microchips through a replica molding technology was achieved in a low-cost setting within 2.5 h. Subsequently, major manufacturing conditions were optimized to acquire well-resolved microfluidic molds, and the replicated microchips were validated to be of good performance. A T-junction channel microchip was fabricated by using a dry film master to generate water droplets of uniform target size. Meanwhile, a gated injection of fluorescein sodium and a contactless conductivity detection of Na⁺ were both performed in a crosslink channel microchip via capillary electrophoresis, in other words, this fast prototyping and fabrication method would be an efficient, economical way to embody structural design into microfluidic chips for various applications.     

A silicon rectangular micro-orifice for gas flow measurement at moderate Reynolds numbers: design,fabrication and flow analyses

This work describes a micro-flowmeter for moderate flow rates of gases based on a differential pressure measurement. The micro-flowmeters consist of a microfabricated silicon–glass rectangular micro-orifice plate, with external pressure measurement. We experimentally evaluate the effects of geometrics parameters, Reynolds number and compressibility on the discharge coefficient. The paper examines a series of 13 rectangular micro-orifice sizes, with orifice hydraulic diameters ranging from 115 to 362 µm. The behavior of the discharge coefficient is presented for orifice Reynolds numbers ranging from 200 to 18000. Agreement is shown between the experimental and numerical results of the discharge coefficient. The micro-flowmeters measure moderate flow of air ranging from 1 to 106 mg/s. This demonstration implements a design method of micro-flowmeters that can be used in a broad range of microfluidic applications, such as microreactors and power MEMS.