Development of Microdroplet Generation Method for Organic Solvents Used in Chemical Synthesis

Recently, chemical operations with microfluidic devices, especially droplet-based operations, have attracted considerable attention because they can provide an isolated small-volume reaction field. However, analysis of these operations has been limited mostly to aqueous-phase reactions in water droplets due to device material restrictions. In this study, we have successfully demonstrated droplet formation of five common organic solvents frequently used in chemical synthesis by using a simple silicon/glass-based microfluidic device. When an immiscible liquid with surfactant was used as the continuous phase, the organic solvent formed droplets similar to water-in-oil droplets in the device. In contrast to conventional microfluidic devices composed of resins, which are susceptible to swelling in organic solvents, the developed microfluidic device did not undergo swelling owing to the high chemical resistance of the constituent materials. Therefore, the device has potential applications for various chemical reactions involving organic solvents. Furthermore, this droplet generation device enabled control of droplet size by adjusting the liquid flow rate. The droplet generation method proposed in this work will contribute to the study of organic reactions in microdroplets and will be useful for evaluating scaling effects in various chemical reactions.     

A simplified PDMS microfluidic device with a built-in suction actuator for rapid production of monodisperse water-in-oil droplets

We previously established an automatic droplet-creation technique that only required air evacuation of a PDMS microfluidic device prior to use. Although the rate of droplet production with this technique was originally slow (∼10 droplets per second), this was greatly improved (∼470 droplets per second) in our recent study by remodeling the original device configuration. This improvement was realized by the addition of a degassed PDMS layer with a large surface area-to-volume ratio that served as a powerful vacuum generator. However, the incorporation of the additional PDMS layer (which was separate from the microfluidic PDMS layer itself) into the device required reversible bonding of five different layers. In the current study, we aimed to simplify the device architecture by reducing the number of constituent layers for enhancing usability of this microfluidic droplet generator while retaining its rapid production rate. The new device consisted of three layers. This comprised a degassed PDMS slab with microfluidic channels on one surface and tens of thousands of vacuum-generating micropillars on the other surface, which was simply sandwiched by PMMA layers. Despite its simplified configuration, this new device created monodisperse droplets at an even faster rate (>1000 droplets per second).     

Generation of Water-In-Oil-In-Water (W/O/W) Double Emulsions by Microfluidics

Novel compartment microparticles prepared with double emulsion droplets as templates provide a protected internal space for material encapsulation. The effect of three-phase flow rate on the micro-droplet generation of double emulsion mechanism is available for reference to produce precise size and highly monodisperse particles. The influence of three-phase flow rate on the formation mode and size of the emulsion droplets is investigated by combination of experiment and numerical simulation. The size of compound droplets decreases and frequency increases with the increasing outer fluid flow rate. The monodispersity of the double emulsion reduces due to transition from dripping to narrowing jetting regime. Outer droplet size increases with the increasing flow rate of the middle fluid, whereas inner droplet size is the opposite. The frequency increases and then stabilizes, which leads to a widening regime. When Q₂/Q₁ > 6, the multi-core type double emulsion droplets are produced. Droplet coalescence occurs when surfactants is not involved. As Q₁ increases, there is an increasing tendency for inner drop size. The outer drop size is proportional to the sum of the inner and middle flow rate, and that is irrelevant to Q₁/Q₂. For drop size, the ratio of core-shell and internal structure is precisely controlled by adjusting three-phase flow rate respectively.     

Highly productive droplet formation by anisotropic elongation of a thread flow in a microchannel

We developed a microfluidic device to form monodisperse droplets with high productivity by anisotropic elongation of a thread flow, defined as a threadlike flow of a dispersed liquid phase in a flow of an immiscible, continuous liquid phase. The thread flow was anisotropically elongated in the depth direction in a straight microchannel with a step, where the microchannel depth changed. Consequently, the elongated thread flow was given capillary instability (Rayleigh-Plateau instability) and was continuously transformed into monodisperse droplets at the downstream area of the step in the microchannel. We examined the effects of the flow rates of the dispersed phase and the continuous phase on the droplet formation behavior, including the droplet diameter and droplet formation frequency. The droplet diameter increased as the fraction of the dispersed-phase flow rate relative to the total flow rate increased and was independent of the total flow rate. The droplet formation frequency proportionally increased with the total flow rate at a constant dispersed-phase flow rate fraction. These results are explained in terms of a mechanism similar to that of droplet formation from a cylindrical liquid thread flow by Rayleigh-Plateau instability. The microfluidic device described was capable of forming monodisperse droplets with a 160-microm average diameter and 3-microm standard deviation at a droplet formation frequency of 350 droplets per second from a single thread flow. The highest total flow rate achieved was 6 mL/h using the present device composed of a straight microchannel with a step. We also demonstrated parallel droplet formation by anisotropic elongation of multiple thread flows; the process was applied to form W/O and O/W droplets. The highly productive droplet formation process presented in this study is expected to be useful for future industrial applications.     

Parallel synchronization of two trains of droplets using a railroad-like channel network

We present a simple method of water-in-oil droplet synchronization in a railroad-like channel network. The network consisted of a top channel, a bottom channel, and ladder-like channels interconnected between the two main channels. The presence of the pressure difference between the top and bottom channels resulted in the crossflow of carrier oil through the ladder network until the pressure in each channel was balanced automatically. The proposed model and method proved the feasibility of the parallel synchronization of two trains of droplets with up to 95% synchronization efficiency. Physical parameters that could improve the efficiency were investigated with the systematic variation of the droplet length and droplet generation frequency by controlling the flow rate in each channel. Under a subtle difference in the generation frequency, an unmatched droplet sandwiched between two matched droplets in the ladder network was switched and synchronized in turn. For perfect one-to-one droplet synchronization, the droplet length and the droplet generation frequency needed to be the same for both the top and bottom channels. In addition, one-to-multiple droplet synchronization was demonstrated by matching the product of the droplet length and the droplet generation frequency for both the top and bottom channels. The proposed method provides a simple unit operation for parallel synchronization of the trains of droplets that can be easily integrated with the conventional continuous-flow droplet-based microfluidic platform.     

Mastering a double emulsion in a simple co-flow microfluidic to generate complex polymersomes

We show that the production and the geometrical shape of complex polymersomes can be predicted by varying the flow rates of a simple microdevice using an empirical law which predicts the droplet size. This device is constituted of fused silica capillaries associated with adjusted tubing sleeves and T-junctions. Studying the effect of several experimental parameters, double emulsions containing a controlled number of droplets were fabricated. First, this study examines the stability of a jet in a simple confined microfluidic system, probing the conditions required for droplets production. Then, multicompartmental polymersomes were formed, controlling flow velocities. In this work, poly(dimethylsiloxane)-graft-poly(ethylene oxide) (PDMS-g-PEO) and poly(butadiene)-block-poly(ethyleneoxide) (PBut-b-PEO) amphiphilic copolymers were used and dissolved in chloroform/cyclohexane mixture. The ratio of these two solvents was adjusted in order to stabilize the double emulsion formation. The aqueous suspension contained poly(vinyl alcohol) (PVA), limiting the coalescence of the droplets. This work constitutes major progress in the control of double emulsion formation in microfluidic devices and shows that complex structures can be obtained using such a process.     

High-volume production of single and compound emulsions in a microfluidic parallelization arrangement coupled with coaxial annular world-to-chip interfaces

This study describes a microfluidic platform with coaxial annular world-to-chip interfaces for high-throughput production of single and compound emulsion droplets, having controlled sizes and internal compositions. The production module consists of two distinct elements: a planar square chip on which many copies of a microfluidic droplet generator (MFDG) are arranged circularly, and a cubic supporting module with coaxial annular channels for supplying fluids evenly to the inlets of the mounted chip, assembled from blocks with cylinders and holes. Three-dimensional flow was simulated to evaluate the distribution of flow velocity in the coaxial multiple annular channels. By coupling a 1.5 cm × 1.5 cm microfluidic chip with parallelized 144 MFDGs and a supporting module with two annular channels, for example, we could produce simple oil-in-water (O/W) emulsion droplets having a mean diameter of 90.7 μm and a coefficient of variation (CV) of 2.2% at a throughput of 180.0 mL h(-1). Furthermore, we successfully demonstrated high-throughput production of Janus droplets, double emulsions and triple emulsions, by coupling 1.5 cm × 1.5 cm - 4.5 cm × 4.5 cm microfluidic chips with parallelized 32-128 MFDGs of various geometries and supporting modules with 3-4 annular channels.     

Efficient Generation of Microdroplets Using Tail Breakup Induced with Multi-Branch Channels

In recent years, research on the application of microdroplets in the fields of biotechnology and chemistry has made remarkable progress, but the technology for the stable generation of single-micrometer-scale microdroplets has not yet been established. In this paper, we developed an efficient and stable single-micrometer-scale droplet generation device based on the fragmentation of droplet tails, called “tail thread mode”, that appears under moderate flow conditions. This method can efficiently encapsulate microbeads that mimic cells and chemical products in passively generated single-micrometer-scale microdroplets. The device has a simple 2D structure; a T-junction is used for droplet generation; and in the downstream, multi-branch channels are designed for droplet deformation into the tail. Several 1–2 µm droplets were successfully produced by the tail’s fragmentation; this continuous splitting was induced by the branch channels. We examined a wide range of experimental conditions and found the optimal flow rate condition can be reduced to one-tenth compared to the conventional tip-streaming method. A mold was fabricated by simple soft lithography, and a polydimethylsiloxane (PDMS) device was fabricated using the mold. Based on the 15 patterns of experimental conditions and the results, the key factors for the generation of microdroplets in this device were examined. In the most efficient condition, 61.1% of the total droplets generated were smaller than 2 μm.     

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.     

Pneumatic handling of droplets on-demand on a microfluidic device for seamless processing of reaction and electrophoretic separation

Sequential operations of pre-separation reaction process by picoliter droplets and following electrophoretic separation process were realized in a single microfluidic device with pneumatic handling of liquid. The developed device consists of a fluidic chip made of PDMS, an electrode substrate, and a temperature control substrate on which thin film heater/sensor structures are fabricated. Liquid handling, including introduction of liquid samples, droplet generation, and merging of droplets, was implemented by pneumatic manipulation through microcapillary vent structures, allowing air to pass and stop liquid flow. Since the pneumatic manipulations are conducted in a fully automated manner by using a programmable air pressure control system, the user simply has to load liquid samples on each liquid port of the device. Droplets of 420 pL were generated with an accuracy of ± 2 pL by applying droplet generation pressure in the range of 40-100 kPa. As a demonstration, a binding reaction of a 15 mer ssDNA with a peptide nucleic acid oligomer used as an oligoprobe followed by denaturing electrophoresis to discriminate a single-base substitution was performed within 1.5 min. By exploiting the droplet-on-demand capability of the device, the influence of various factors, such as reaction time, mixing ratio and droplet configurations on the ssDNA-peptide nucleic acid binding reaction in the droplet-based process, was studied toward realization of a rapid detection method to discriminate rapid single-base substitution.     

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.     

Preparation of Poly(vinyl alcohol) Microspheres Based on Droplet Microfluidic Technology

A new method for preparing poly (vinyl alcohol) (PVA) microspheres was developed by using droplet microfluidic technology. In the microfluidic chip, a large number of uniform, monodispersed PVA droplets were prepared quickly and continuously by using droplet formation technology, and the droplet preparation speed reached 7 per second. The size of the PVA droplets could be controlled by changing the injection flow rate of the two-phase fluid and the width of microfluidic channel. Then the PVA microspheres were formed by physical crosslinking. This method has high preparation efficiency and good monodispersity of the obtained microspheres. Moreover, the process does not require the incorporation of chemical crosslinking agents, avoiding interference with the inclusion material, and is well suited for applications such as drug carrier.     

Alternating droplet generation and controlled dynamic droplet fusion in microfluidic device for CdS nanoparticle synthesis

A multifunctional and high-efficiency microfluidic device for droplet generation and fusion is presented. Through unique design of the micro-channels, the device is able to alternately generate droplets, generating droplet ratios ranging from 1 ratio 5 to 5 ratio 1, and fuse droplets, enabling precise chemical reactions in several picoliters on a single chip. The controlled fusion is managed by passive control based on the channel geometry and liquid phase flow. The synthesis of CdS nanoparticles utilizing each fused droplet as a microreactor for rapid and efficient mixing of reagents is demonstrated in this paper. Following alternating droplet generation, the channel geometry allows the exclusive fusion of alternate droplets with concomitant rapid mixing and produces supersaturated solution of Cd2+ and S2- ions to form CdS nanoparticles in each fused droplet. The spectroscopic properties of the CdS nanoparticles produced by this method are compared with CdS prepared by bulk mixing.     

Three-dimensional interconnected microporous poly(dimethylsiloxane) microfluidic devices

This technical note presents a fabrication method and applications of three-dimensional (3D) interconnected microporous poly(dimethylsiloxane) (PDMS) microfluidic devices. Based on soft lithography, the microporous PDMS microfluidic devices were fabricated by molding a mixture of PDMS pre-polymer and sugar particles in a microstructured mold. After curing and demolding, the sugar particles were dissolved and washed away from the microstructured PDMS replica revealing 3D interconnected microporous structures. Other than introducing microporous structures into the PDMS replica, different sizes of sugar particles can be used to alter the surface wettability of the microporous PDMS replica. Oxygen plasma assisted bonding was used to enclose the microstructured microporous PDMS replica using a non-porous PDMS with inlet and outlet holes. A gas absorption reaction using carbon dioxide (CO(2)) gas acidified water was used to demonstrate the advantages and potential applications of the microporous PDMS microfluidic devices. We demonstrated that the acidification rate in the microporous PDMS microfluidic device was approximately 10 times faster than the non-porous PDMS microfluidic device under similar experimental conditions. The microporous PDMS microfluidic devices can also be used in cell culture applications where gas perfusion can improve cell survival and functions.     

Nucleation rate measurement of colloidal crystallization using microfluidic emulsion droplets

An emulsion crystallization method has been demonstrated to measure the nucleation rate of a thermoresponsive colloidal poly-N-isopropylacrylamide (PNIPAM) system. The colloidal PNIPAM suspension was injected into a microfluidic flow-focusing device to generate monodispersed droplets in oil. The temperature was controlled to fine tune the volume fraction of the PNIPAM particles, and the microfluidic flow rate was varied to change the droplet sizes, thus altering the nucleation volume. Using independent droplets, we can isolate the nucleation events to eliminate the interactions among crystallites that existed in bulk or large droplet systems. Therefore, we were able to carry out accurate nucleation rate measurements of colloidal crystals. This emulsion crystallization method is promising for bridging the gap among theories, simulations, and experiments for nucleation kinetics studies.     

Static droplet array for the synthesis of nonspherical microparticles

We reported a manually operated static droplet array (SDA)-based device for the synthesis of nonspherical microparticles with different shapes. The improved SDA structure and reversible bonding between poly(dimethylsiloxane) (PDMS) were used in the device for the large-scale synthesis and rapid extraction of nonspherical microparticles. To understand the device physics, the effects of flow rate, SDA well size, and shape on droplet generation performances were explored. The results indicated that droplet generation in SDA structures was insensitive to the flow rate, and monodisperse droplets were generated by the SDA-based device through manually pushing the syringe. Finally, we integrated four kinds of SDA structures in one device and successfully realized the synthesis and extraction of nonspherical microparticles with different shapes and materials. Our SDA-based device offers numerous advantages, such as simple manual operation, low equipment cost, controllable microparticle shapes and sizes, and large-scale production. Thus, it holds the potential to be used as a flexible tool for the production of nonspherical microparticles.     

Microfluidic formation of highly monodispersed multiple cored droplets using needle-based system in parallel mode

Scale-up in droplet microfluidics achieved by increasing the number of devices running in parallel or increasing the droplet makers in the same device can compromise the narrow droplet-size distribution, or requires high fabrication cost, when glass- or polymer-based microdevices are used. This paper reports a novel way using parallelization of needle-based microfluidic systems to form highly monodispersed droplets with enhanced production rates yet in cost-effective way, even when forming higher order emulsions with complex inner structure. Parallelization of multiple needle-based devices could be realized by applying commercially available two-way connecters and 3D-printed four-way connectors. The production rates of droplets could be enhanced around fourfold (over 660 droplets/min) to eightfold (over 1300 droplets/min) by two-way connecters and four-way connectors, respectively, for the production of the same kind of droplets than a single droplet maker (160 droplets/min). Additionally, parallelization of four-needle sets with each needle specification ranging from 34G to 20G allows for simultaneous generation of four groups of PDMS microdroplets with each group having distinct size yet high monodispersity (CV < 3%). Up to six cores can be encapsulated in double emulsion using two parallelly connected devices via tuning the capillary number of middle phase in a range of 1.31 × 10⁻⁴ to 4.64 × 10⁻⁴. This study leads to enhanced production yields of droplets and enables the formation of groups of droplets simultaneously to meet extensive needs of biomedical and environmental applications, such as microcapsules with variable dosages for drug delivery or drug screening, or microcapsules with wide range of absorbent loadings for water treatment.     

Fabrication of poly(dimethylsiloxane) microfluidic system based on masters directly printed with an office laser printer

Applications of poly(dimethylsiloxane) (PDMS)-based microfluidic systems are more popular nowadays. Previous fabrication methods of the masters for PDMS microchannels require complicated steps and/or special device. In this paper, we demonstrated that the toner printed on the transparency film with the office laser printer (1200 dpi) can be used as the positive relief of the masters. The transparency film was printed in two steps in order to obtain the same printing quality for the crossed lines. With the laser-printed master, the depth of the fabricated PDMS microchannels was ca. 10 microm and the smallest width was ca. 60 microm. Surface characteristics of the PDMS/PDMS microchannels were performed with SEM. Their electrokinetic properties were investigated by the aids of the measurement of electroosmotic flow (EOF) and the Ohm's curve. Using the PDMS/PDMS microchip CE systems, electroactive biological molecules and non-electroactive inorganic ions were well separated, respectively. This simple approach could make it easy to carry out the studies of PDMS microfluidic systems in more general labs without special devices.     

A single-cell encapsulation method based on a microfluidic multi-step droplet splitting system

Single cell analysis is of great significance to understand the physiological activity of organisms.Microfluidic droplet is an ideal analytical platform for single-cell analysis. We developed a microfluidic droplet splitting system integrated with a flow-focusing structure and multi-step splitting structures to form 8-line droplets and encapsulate single cells in the droplets. Droplet generation frequency reached1021 Hz with the aqueous phase flow rate of 1 m L/min and the oil phase flow rate of 15 mL /min. Relative standard deviation of the droplet size was less than 5% in a single channel, while less than 6% in all the8 channels. The system was used for encapsulating human whole blood cells. A single-cell encapsulation efficiency of 31% was obtained with the blood cell concentration of 2.5× 10~4cells/mL, and the multicellular droplet percentage was only 1.3%. The multi-step droplet splitting system for single cell encapsulation featured simple structure and high throughput.     

An electrochemically driven poly(dimethylsiloxane) microfluidic actuator: oxygen sensing and programmable flows and pH gradients

We describe the fabrication and performance of an integrated microelectrochemical reactor-a design possessing utility for multiple applications that include electrochemical sensing, the generation and manipulation of in-channel microfluidic pH gradients, and fluid actuation and flow. The device architecture is based on a three-electrode electrochemical cell design that incorporates a Pt interdigitated array (IDA) working (WE), a Pt counter (CE), and Ag pseudo-reference (RE) electrodes within a microfluidic network in which the WE is fully immersed in a liquid electrolyte confined in the channels. The microchannels are made from a conventional poly(dimethylsiloxane)(PDMS) elastomer, which serves also as a thin gas-permeable membrane through which gaseous reactants in the external ambient environment are supplied to the working electrode by diffusion. Due to the high permeability of oxygen through PDMS, the microfluidic cell supports significantly (>order of magnitude) higher current densities in the oxygen reduction reaction (ORR) than those measured in conventional (quiescent) electrochemical cells for the same electrode areas. We demonstrate in this work that, when operated at constant potential under mass transport control, the device can be utilized as a membrane-covered oxygen sensor, the response of which can be tuned by varying the thickness of the PDMS membrane. Depending on the experimental conditions under which the electrochemical ORR is performed, the data establish that the device can be operated as both a programmable pH gradient generator and a microfluidic pump.