Fabrication of polymersomes with controllable morphologies through dewetting w/o/w double emulsion droplets

In this work, monodisperse giant polymersomes are fabricated by dewetting of water-in-oil-in-water double emulsion droplets which are assembled by amphiphilic block copolymer molecules in a microfluidic device. The dewetting process can be tuned by solvation between solvent and amphiphilic block copolymer to get polymersomes with controllable morphology. Good solvent(chloroform and toluene) hinders dewetting process of double emulsion droplets and gets acornlike polymersomes or patched polymersomes. On the other hand, poor solvent(hexane) accelerates the dewetting process and achieves complete separation of inner water phase from oil phase to form complete bilayer polymersomes. In addition, twin polymersomes with bilayer membrane structure are formed by this facile method. The formation mechanism for different polymersomes is discussed in detail.     

Dewetting instability during the formation of polymersomes from block-copolymer-stabilized double emulsions

We investigate the formation of polymer vesicles, or polymersomes, of polystyrene-block-poly(ethylene oxide) diblock copolymers using double emulsion droplets of controlled architecture as templates. To engineer the structure of the polymersomes, it is important to consider the concentration of diblock copolymer in the middle phase of the double emulsion. We describe how the presence of excess polymer can induce a transition from complete wetting to partial wetting of the middle phase, resulting in polymer shells with inhomogeneous thicknesses.     

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.     

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.     

The Effect of Stabilizer on the Mechanical Response of Double‐Emulsion‐Templated Polymersomes

Recent studies have shown that polymersomes templated by microfluidic double‐emulsion possess several advantages such as high monodispersity and encapsulation efficiency compared with those generated based on thin‐film rehydration and electroformation. Stabilizers, including bovine serum albumin (BSA) and polyvinyl alcohol (PVA), have been used to enhance the formation and stability of double emulsions that are used as templates for the generation of polymersomes. In this work, the effect of stabilizers on the mechanical response of double‐emulsion‐templated polymersomes using micropipette aspiration is investigated. It is demonstrated that the existence of stabilizers results in the inelastic response in poly­mersomes in the early stage of solvent removal. However, aged polymersomes that have little residual solvent show elastic behavior. Polymersomes prepared from PVA‐stabilized double emulsions have noticeably lower area expansion moduli than polymersomes prepared from stabilizer‐free and BSA‐stabilized double emulsions, suggesting that PVA is incorporated in the bilayer membrane of polymersomes.

     

Hierarchical Porous Polymer Beads Prepared by Polymerization-induced Phase Separation and Emulsion-template in a Microfluidic Device

Porous polymer beads（PPBs） containing hierarchical bimodal pore structure with gigapores and meso-macropores were prepared by polymerization-induced phase separation（PIPS） and emulsion-template technique in a glass capillary microfluidic device（GCMD）. Fabrication procedure involved the preparation of water-in-oil emulsion by emulsifying aqueous solution into the monomer solution that contains porogen. The emulsion was added into the GCMD to fabricate the（water-in-oil）-in-water double emulsion droplets. The flow rate of the carrier continuous phase strongly influenced the formation mechanism and size of droplets. Formation mechanism transformed from dripping to jetting and size of droplets decreased from 550 μm to 250 μm with the increase in flow rate of the carrier continuous phase. The prepared droplets were initiated for polymerization by on-line UV-irradiation to form PPBs. The meso-macropores in these beads were generated by PIPS because of the presence of porogen and gigapores obtained from the emulsion-template. The pore morphology and pore size distribution of the PPBs were investigated extensively by scanning electron microscopy and mercury intrusion porosimetry（MIP）. New pore morphology was formed at the edge of the beads different from traditional theory because of different osmolarities between the water phase of the emulsion and the carrier continuous phase. The morphology and proportion of bimodal pore structure can be tuned by changing the kind and amount of porogen.     

Synthesis of titania-silica core-shell microspheres via a controlled interface reaction in a microfluidic device

In this work, we describe a novel, simple microfluidic method for fabricating titania-silica core-shell microspheres. Uniform droplets of silica sol were dispersed into an oil phase containing tetrabutyl titanate via a coaxial microfluidic device. The titanium alkoxide hydrolyzed at the water-oil interface after the formation of the aqueous droplets. A gel shell containing the titanium hydroxide formed around the droplets, and the titania-silica core-shell microspheres were obtained after calcinations. The X-ray diffraction results show that titania coatings crystallized into a pure anatase structure. The scanning electron microscopy and energy-dispersive spectrometry characterization shows that the microspheres are monodispersed with uniform titania coating on the surface. The dispersity and size of the microspheres could easily be controlled by changing the microfluidic flow parameters. The titania content on the surface could be adjusted in the large range of 1.0-98.0 mol % by varying the continuous phase composition and the reaction time, and the structures of the core-shell microshperes could also be controlled.     

Can polymersomes form colloidosomes?

Hydroxy-functionalized polymersomes (or block copolymer vesicles) were prepared via a facile one-pot RAFT aqueous dispersion polymerization protocol and evaluated as Pickering emulsifiers for the stabilization of emulsions of n-dodecane emulsion droplets in water. Linear polymersomes produced polydisperse oil droplets with diameters of ～50 μm regardless of the polymersome concentration in the aqueous phase. Introducing an oil-soluble polymeric diisocyanate cross-linker into the oil phase prior to homogenization led to block copolymer microcapsules, as expected. However, TEM inspection of these microcapsules after an alcohol challenge revealed no evidence for polymersomes, suggesting these delicate nanostructures do not survive the high-shear emulsification process. Thus the emulsion droplets are stabilized by individual diblock copolymer chains, rather than polymersomes. Cross-linked polymersomes (prepared by the addition of ethylene glycol dimethacrylate as a third comonomer) also formed stable n-dodecane-in-water Pickering emulsions, as judged by optical and fluorescence microscopy. However, in this case the droplet diameter varied from 50 to 250 μm depending on the aqueous polymersome concentration. Moreover, diisocyanate cross-linking at the oil/water interface led to the formation of well-defined colloidosomes, as judged by TEM studies. Thus polymersomes can indeed stabilize colloidosomes, provided that they are sufficiently cross-linked to survive emulsification.     

Formation and characterization of polymersomes made by a solvent injection method

In this article a solvent injection method is described for vesicle formation using poly(butadiene)‐ b‐poly(acrylic acid) diblock copolymers as the amphiphilic molecules. Vesicles composed of polymer bilayers are commonly referred to as polymersomes. Solvent injection is shown to be a rapid method for polymersome formation suitable to make large volumes of polymersome solution. The method can be manipulated to obtain a wide range of vesicle sizes depending on the polymer concentration and preparation conditions. Polymersome solutions in this study are characterized using dynamic light scattering (DLS), fluorescent microscopy, and electron microscopy. Polymersome sizes range from tens of nanometers to several microns. The membrane thickness of smaller polymersomes is found to lie between 14–20 nm. Larger polymersomes are found to have somewhat thicker membranes. The procedure involves the addition of polymers dissolved in an organic solvent to a stirred aqueous solution. The formation of polymersomes by this method is proposed to be governed by the limited mutual solubility of the two solvents and the simultaneous diffusion of solvent and water out of and in to initially formed organic solvent droplets. Copyright © 2007 John Wiley & Sons, Ltd.     

Microfluidic Formation of Membrane‐Free Aqueous Coacervate Droplets in Water

We report on the formation of coacervate droplets from poly(diallyldimethylammonium chloride) with either adenosine triphosphate or carboxymethyl‐dextran using a microfluidic flow‐focusing system. The formed droplets exhibit improved stability and narrower size distributions for both coacervate compositions when compared to the conventional vortex dispersion techniques. We also demonstrate the use of two parallel flow‐focusing channels for the simultaneous formation and co‐location of two distinct populations of coacervate droplets containing different DNA oligonucleotides, and that the populations can coexist in close proximity up to 48 h without detectable exchange of genetic information. Our results show that the observed improvements in droplet stability and size distribution may be scaled with ease. In addition, the ability to encapsulate different materials into coacervate droplets using a microfluidic channel structure allows for their use as cell‐mimicking compartments.     

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.     

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.     

Controllable preparation of nanoparticle-coated chitosan microspheres in a co-axial microfluidic device

In this work, we describe a novel and simple microfluidic method for fabricating nanoparticle-coated chitosan microspheres. Uniform droplets of aqueous chitosan solution were dispersed into an oil phase containing partially hydrophilic nanoparticles via a co-axial microfluidic device. Recirculating flow in the continuous phase in the area between drops enhanced mixing and allowed the nanoparticles to coat the surface of the droplets as they passed through the channel. The chitosan droplets were then crosslinked with glutaraldehyde and nanoparticle-coated microspheres were obtained. SEM characterization shows that the microspheres are monodispersed with uniform nanoparticle distribution on the surface. The dispersity, size and composition of the microspheres could all easily be controlled by changing the microfluidic flow parameters and three different types of nanoparticles were successfully used to synthesize hybrid microspheres to demonstrate the method's versatility.     

Biopolymer microparticle and nanoparticle formation within a microfluidic device

This paper reports a novel microfluidic method for the production of cross-linked alginate microparticles and nanoparticles. We describe a continuous process relying on both thermodynamic and hydrodynamic factors to form microdroplets. A rapid cross-linking reaction thereafter allows solidification of the polymer droplets either within the microfluidic device or "off-chip" to form alginate micro- and nanoparticles. Monodisperse droplets are generated by extruding an aqueous alginate solution using an axisymmetric flow-focusing design. As they flow downstream in the channel, due to water and the continuous phase being partially miscible, the water diffuses very slowly out of the polymeric droplets into the transport fluid, which causes the shrinkage of the drops and the condensation of the polymer phase. The resulting size of the solid particles depends on the polymer concentration and the ensuing balance between the kinetics of the cross-linking reaction and the volume loss due to solvent diffusion. This work details both a single-step microfluidic technique for the formation of alginate microparticles of sizes ranging from 1 to 50 microm via near-equilibrium solvent diffusion within a microfluidic device and thereafter a two-step method, which was shown to generate biopolymer nanoparticles of sizes ranging from 10 to 300 nm. These novel methodologies are extremely flexible and can be extended to the preparation of micro- and nanoparticles from a wide range of single or mixed synthetic and biologically derived polymers.     

Numerical simulation of double emulsion formation in cross-junctional flow-focusing microfluidic device using Lattice Boltzmann method

This paper presents an analysis of double emulsion formation through a cross junctional flow-focusing microchannel which uses an improved color gradient lattice Boltzmann method. The model with the potential to simulate a ternary system of immiscible fluids was employed. Two double emulsion formation regimes, one-step and two-step, were simulated. The effect of the inner flow rate, as well as inner and outer flow viscosity was investigated on double emulsion formation. The inner flow rate had a significant influence on the inner liquid jet detachment point in the two-step process. However, the viscosity of the inner and outer fluid considerably affected the double jet detachment point in the one-step formation. The shell thickness of a double emulsion can be adjusted by altering the inner flow rate.     

Micro-droplet detection and characterization using thermal responses

We suggest a novel method to detect droplets and determine the protein content of droplets in microfluidic system using the 3ω method, which is a powerful tool to easily detect thermal response changes with a simple device. By measuring the thermal response of droplets and a carrying flow in real time, water droplets in an oleic acid carrying flow can be detected, and the concentration of bovine serum albumin in droplets can be estimated. This method is expected to increase the practicality and power of droplet-based microfluidic systems.     

Microfluidic large-scale integration on a chip for mass production of monodisperse droplets and particles

In this study, we report the mass production of monodisperse emulsion droplets and particles using microfluidic large-scale integration on a chip. The production module comprises a glass microfluidic chip with planar microfabricated 16-256 droplet-formation units (DFUs) and a palm-sized stainless steel holder having several layers for supplying liquids into the inlets of the mounted chip. By using a module having 128 cross-junctions (i.e., 256 DFUs) arranged circularly on a 4 cm x 4 cm chip, we could produce droplets of photopolymerizable acrylate monomer at a throughput of 320.0 mL h(-1). The product was monodisperse, having a mean diameter of 96.4 microm, with a coefficient of variation (CV) of 1.3%. Subsequent UV polymerization off the module yielded monodisperse acrylic microspheres at a throughput of approximately 0.3 kg h(-1). Another module having 128 co-flow geometries could produce biphasic Janus droplets of black and white segments at 128.0 mL h(-1). The product had a mean diameter of 142.3 microm, with a CV of 3.3%. This co-flow module could also be applied in the mass production of homogeneous monomer droplets.     

Controlled generation of submicron emulsion droplets via highly stable tip-streaming mode in microfluidic devices

Submicron emulsions could be produced via the tip-streaming process in a flow-focusing microfluidic device. In this article, the stability of the liquid cone and thread for tip-streaming mode could be significantly improved by employing a three-dimensional flow-focusing device, in which the hydraulic resistance was adjusted by modulating the channel heights in the flow focusing area, orifice, downstream and dispersed phase inlet channel. The pressure range for tip-streaming mode was enlarged significantly compared with two-dimensional flow-focusing devices. Therefore, monodisperse emulsions were produced under this tip-streaming mode for as long as 48 hours. Furthermore, we could control the size of emulsion drops by changing the pressure ratio in three-dimensional flow-focusing devices while the liquid cone was easily retracted during the adjustment of pressure ratio in two-dimensional flow-focusing devices. Furthermore, using the uniform submicron emulsion droplets as confining templates, polyethylene glycol (PEG) particles were produced with a narrow size distribution at the sub-micrometre scale. In addition, magnetic nanoparticles were added to the emulsion for magnetic PEG particles, which can respond to magnetic field and would be biocompatible.     

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.     

Production of porous suspension polymer beads with a narrow size distribution using a cross-flow membrane and a continuous tubular reactor

The work described focuses on a two-stage process for the production of large porous suspension polymer beads of defined particle size and narrow size distribution. Emulsification has been performed using purpose built cross-flow membrane equipment, which allows controlled production of large emulsion droplets with a much narrower size distribution. The work described compares the production of large emulsion droplets of monomer prepared both by agitation and using a cross-flow membrane. The effects of variations in the pore size of the membrane and flow-rates on the size of the emulsion droplets produced are also investigated. The second stage of the process is polymerisation of performed monomer emulsion droplets using a continuous tubular reactor. Samples polymerised using such a method show a narrower size distribution than similar systems polymerised as a batch.