Microfluidic Emulsification Research Articles

Photoresponsive microfluidic three-phase emulsions for tandem reactions

The long-term objective of tandem reaction is to investigate ‘intelligent’ tandem reaction systems akin to living cells. Here we’ve prepared an ‘intelligent’ droplet microreactor for the tandem reaction using droplet microfluidic technology constructing a photoresponsive microfluidic tandem reaction system. The method leverages a three-phase emulsion’s ‘two membranes and three phases’ structure to segregate incompatible reagents, and introduces a stimulating surfactant to control the structure of this emulsion structure for intelligent reaction sequence control, enablingefficient,orderlytandem reactionwithin the same system. Validating this concept, we applied the droplet microreactor to the condensation-reduction cascade reaction, as envisioned, incompatible condensation and reduction reactions, were coordinated by cycling the UV light on /off for a smooth cascade reaction. Compared to batch reaction, the droplet microreactor showed sixfold catalytic efficiency enhancement and > 99 % reaction selectivity. Our work offers novel insights into realizing intelligent cascade reactions.

Tailoring esophageal tumor spheroids on a chip with inverse opal scaffolds for drug screening

Esophageal cancer (EC) is characterized by high morbidity and mortality, and chemotherapy has become an indispensable means for comprehensive treatment. However, due to the limitation of the effective in vitro disease model, the development of chemotherapeutic agents still faces great challenges. In this paper, we present a novel tumor spheroid on a chip platform based on inverse opal hydrogel scaffolds to screen chemotherapeutic agents for EC treatment. With the microfluidic emulsion approach, the inverse opal hydrogel scaffolds were generated with tunable and organized pores, which could provide spatial confinement for cell growth. Thus, the suspended KYSE-70 cells could successfully form uniform cell spheroids on the inverse opal hydrogel scaffolds. It was demonstrated that the tumor cell spheroids could recapitulate 3D growth patterns in vivo and exhibited higher sensitivity to the chemotherapy agents compared with monolayer cells. Besides, by employing the scaffolds into a microfluidics to construct esophageal tumor on a chip, the device could realize high-throughput tumor cell spheroids generation and drug screening, indicating its promising role in chemotherapy drug development.

Microfluidic Step Emulsification with Parallel Nozzles on a Vertical Slit

Microfluidic Step Emulsification with Parallel Nozzles on a Vertical Slit

Microstructured Porous Capacitive Bio-pressure Sensor Using Droplet-based Microfluidics.

Devices that mimic the functions of human skin are known as "electronic skin," and they must have characteristics like high sensitivity, a wide dynamic range, high spatial homogeneity, cheap cost, wide area easy processing, and the ability to distinguish between diverse external inputs. This study introduces a novel approach, termed microfluidic droplet-based emulsion self-assembly (DMESA), for fabricating 3D microstructured elastomer layers using polydimethylsiloxane (PDMS). The method aims to produce accurate capacitive pressure sensors suitable for electronic skin (e-skin) applications. The DMESA method facilitates the creation of uniform-sized spherical micropores dispersed across a significant area without requiring a template, ensuring excellent spatial homogeneity. Micropore size adjustment, ranging from 100 to 600 μm, allows for customization of pressure sensor sensitivity. The active layer of the capacitive pressure sensor is formed by the three-dimensional elastomer itself. Experimental results demonstrate the outstanding performance of the DMESA approach. It offers simplicity in processing, the ability to adjust performance parameters, excellent spatial homogeneity, and the capability to differentiate varied inputs. Capacitive pressure sensors fabricated using this method exhibit high sensitivity and dynamic amplitude, making them promising candidates for various e-skin applications. The DMESA method presents a highly promising solution for fabricating 3D microstructured elastomer layers for capacitive pressure sensors in e-skin technology. Its simplicity, performance adjustability, spatial homogeneity, and sensitivity to different inputs make it suitable for a wide range of electronic skin applications.

Droplet size dependency and spatial heterogeneity of lipid oxidation in whey protein isolate-stabilized emulsions

Spatiotemporal assessment of lipid and protein oxidation is key for understanding quality deterioration in emulsified food products containing polyunsaturated fatty acids. In this work, we first mechanistically validated the use of the lipid oxidation-sensitive fluorophore BODIPY 665/676 as a semi-quantitative marker for local peroxyl radical formation. Next, we assessed the impact of microfluidic and colloid mill emulsification (respectively producing mono- and polydisperse droplets) on local protein and lipid oxidation kinetics in whey protein isolate (WPI)-stabilized emulsions. We further used BODIPY 581/591 C11 and CAMPO-AFDye 647 as colocalisation markers for lipid and protein oxidation. The polydisperse emulsions showed an inverse relation between droplet size and lipid oxidation rate. Further, we observed less protein and lipid oxidation occurring in similar sized droplets in monodisperse emulsions. This observation was linked to more heterogeneous protein packing at the droplet surface during colloid mill emulsification, resulting in larger inter-droplet heterogeneity in both protein and lipid oxidation. Our findings indicate the critical roles of emulsification methods and droplet sizes in understanding and managing lipid oxidation.

Unravelling the effect of droplet size on lipid oxidation in O/W emulsions by using microfluidics

Lipid oxidation in emulsions is hypothesised to increase with decreasing droplet size, as this increases the specific oil–water interfacial area, where lipid oxidation is expected to be initiated. In literature, however, contradictory results have been reported, which can be caused by confounding factors such as the oil droplet polydispersity and the distribution of components between the available phases. In this work, monodisperse surfactant-stabilised emulsions with highly controlled droplet sizes of 4.7, 9.1, and 26 µm were produced by microfluidic emulsification. We show that lipid oxidation increases with decreasing droplet size, which we ascribe to the increased contact area between lipids and continuous phase prooxidants. Besides, a significant amount of oxygen was consumed by oxidation of the surfactant itself (Tween 20), an effect that also increased with decreasing droplet size. These insights substantiate the importance of controlling droplet size for improving the oxidative stability of emulsions.

Programmable Enzymatic Reaction Network in Artificial Cell-Like Polymersomes.

The ability to precisely control in vitro enzymatic reactions in synthetic cells plays a crucial role in the bottom-up design of artificial cell models that can recapitulate the key cellular features and functions such as metabolism. However, integration of enzymatic reactions has been limited to bulk or microfluidic emulsions without a membrane, lacking the ability to design more sophisticated higher-order artificial cell communities for reconstituting spatiotemporal biological information at multiple length scales. Herein, droplet microfluidics is utilized to synthesize artificial cell-like polymersomes with distinct molecular permeability for spatiotemporal control of enzymatic reactions driven by external signals and fuels. The presence of a competing reverse enzymatic reaction that depletes the active substrates is shown to enable demonstration of fuel-driven formation of sub-microcompartments within polymersomes as well as realization of out-of-equilibrium systems. In addition, the different permeability characteristics of polymersome membranes are exploited to successfully construct a programmable enzymatic reaction network that mimics cellular communication within a heterogeneous cell community through selective molecular transport.

Effects of Rheological Properties of Non-Newtonian Middle Fluids on the Formation Dynamics of Double Emulsions in Coflowing Microfluidic Emulsification

Effects of Rheological Properties of Non-Newtonian Middle Fluids on the Formation Dynamics of Double Emulsions in Coflowing Microfluidic Emulsification

In vitro vascularized liver tumor model based on a microfluidic inverse opal scaffold for immune cell recruitment investigation.

Liver cancer, characterized as a kind of malignant tumor within the digestive system, poses great health harm, and immune escape stands out as an important reason for its occurrence and development. Chemokines, pivotal in guiding immune cells' migration, is necessary to initiate and deliver an effective anti-tumor immune response. Therefore, understanding the chemotactic environment and identifying chemokines that regulate recruitment of immune cells to the tumor microenvironment (TME) are critical to improve current immunotherapy interventions. Herein, we report a well-defined inverse opal scaffold generated with a microfluidic emulsion template for the construction of a vascularized liver tumor model, offering insights into immune cells' recruitment. Due to the excellent 3D porous morphology of the inverse opal scaffold, human hepatocellular carcinoma cells can aggregate in the pores of the scaffold to form uniform multicellular tumor spheroids. More attractively, the vascularized liver tumor model can be achieved by constructing a 3D co-culture system involving endothelial cells and hepatocellular carcinoma cells. The results demonstrate that the 3D co-cultured tumor cells increase the neutrophil chemokines remarkably and recruit neutrophils to tumor tissues, then promote tumor progression. This approach opens a feasible avenue for realizing a vascularized liver tumor model with a reliable immune microenvironment close to that of a solid tumor of liver cancer.

Light‐Induced Virtual Electrodes for Microfluidic Droplet Electro‐Coalescence

AbstractElectro‐coalescence is the fusion phenomenon between a pair or more microfluidic droplets that are immersed in an immiscible medium under an electric field. This technique is frequently used to merge confined droplets in surfactant‐stabilized microfluidic emulsions using local electric fields. Despite the necessity of miniaturized electrodes, this method has proven highly successful in microfluidics and lab‐on‐a‐chip applications. Miniaturized electrodes severely curtail the spatial and temporal flexibility of the electric potential, thus hindering real‐time and flexible operation and leading to high production costs. The current study addresses this problem with reconfigurable electric field potential by light‐driven functional virtual electrodes. These electrodes are light‐induced on a non‐centrosymmetric ferroelectric photovoltaic crystal placed below a microfluidic droplet channel. The photovoltaic effect in the crystal is responsible for the space charge distributions that act as virtual electrodes, whose evanescent field is screened by free charges into the two liquids inside the channel. A numerical model is developed to describe the evolution of the evanescent electric field causing electro‐coalescence. Based on this prediction, two coalescence processes occur at two different timescales and with different numbers of droplets involved. Controlled exposure time modulation allows either rapid on‐demand coalescence of droplet pairs or breakup of the entire emulsion.

Breadmaking-Inspired Antioxidant Porous Yeast Microcarriers for Stem Cell Delivery in Diabetic Wound Treatment.

Stem cell-based therapies have exhibited significant promise in the treatment of diabetic ulcers (DU). Nevertheless, enhancing the survival rate and functionality of transplanted stem cells poses a substantial challenge. In this study, inspired by the breadmaking process, yeast microcarriers (YMC) are devised as vehicles for stem cells to address these challenges. The fabrication of YMC involves the amalgamation of microfluidic emulsification with yeast-mediated fermentation, yielding microcarriers with outstanding biocompatibility, high porosity, and antioxidant activity. Adipose-derived stem cells (ADSCs) seeded onto YMC display remarkable cell viability and retain their cellular functions effectively. Additionally, YMC boast a rich glutathione content and exhibit remarkable ROS scavenging ability, thus shielding the ADSCs from oxidative stress. In vivo experiments further substantiate that ADSC@YMC implementation significantly lowered ROS levels in diabetic wounds, resulting in enhanced stem cell retention and improved angiogenesis, collagen deposition, and tissue regeneration. These results highlight the potential of ADSC@YMC as a promising platform for delivering stem cell in the treatment of diabetic wounds.

Pitaya-inspired microcarrier/hydrogel composite for chronic wound healing: One-pot preparation, tailorable structures, and versatile functionalization

Development of hydrogel-based constructs that integrate active substances appears to be a promising direction for treating chronic wounds. However, their practical applications are not satisfied as expected and limited by cumbersome fabrication processes, high production costs, uncontrolled compositions, and lacking multiple functions into one platform. Herein, inspired by the structure of a pitaya, a microcarrier-embedded hydrogel-based construct is designed via one-pot microfluidic emulsification. The hydrogel matrix like the creamy white flesh of a pitaya is composed of snail mucus and alginate, possessing demanded bio-adhesion, bio-activity, and bio-compatibility. The embedded microcarriers like the small black seeds of a pitaya are composed of mesoporous silica nanoparticles and poly(lactic-co-glycolic acid), serving as an excellent device to control drug delivery. Benefited from the emulsion manipulation ability of microfluidic system, the content ratio of embedded microcarriers in hydrogel matrix, and the respective compositions of embedded microcarriers and hydrogel matrix are all variable to adapt to the individualized treatment. Importantly, the pitaya-inspired microcarrier/hydrogel composite (PMH) can be further functionalized by introducing specific actives into its hierarchical compartments. After encapsulating silver nanoparticles and angiogenic molecules into the hydrogel matrix and embedded microcarriers respectively, both in vitro and in vivo experiments demonstrate that PMH can prevent bacteria invasion, promote cell function, and accelerate wound healing under diabetic conditions with enhanced epidermis regeneration, collagen deposition, blood vessel formation, and skin function recovery. Collectively, taking advantages of its one-pot preparation, tailorable structures, and versatile functionalization, the broad application of PMH in chronic wound management and other biomedical fields is anticipated.

Controlled production of liposomes with novel microfluidic membrane emulsification for application of entrapping hydrophilic and lipophilic drugs

In view of the limitations in the reported methods for the preparation of liposomes, a new microfluidic membrane emulsification device was innovatively designed and fabricated to investigate the effects of prescription factors and process parameters on the formation of liposomes based on a liposome prescription suitable for encapsulating nucleotides. It was shown that precise control of liposome particle size can be achieved by adjusting prescription factors and process parameters, and the obtained liposomes have good stability. In this study, we efficiently encapsulated nicotinamide mononucleotide (NMN) and honokiol to achieve co-delivery and graded release of hydrophilic and hydrophobic drugs, and its inhibitory activity against colon cancer cells was significantly higher than that of the free drug. What’s more, a new high-performance liquid chromatographic method was built up to determine the encapsulation rate of NMN liposomes, which would be useful for the development of NMN drug applications. Notably, we also extended the technique to drug delivery microsphere platform construction to achieve homogeneous and controllable preparation of high-viscosity calcium alginate microspheres. This study is a valuable guide for the construction of drug delivery system platforms and has promising applications.

Experimental evolution of Pseudomonas aeruginosa to colistin in spatially confined microdroplets identifies evolutionary trajectories consistent with adaptation in microaerobic lung environments.

Antibiotic resistance remains one of the great challenges confronting public health in the world today. Individuals with compromised immune systems or underlying health conditions are often at an increased for bacterial infections. Patients with cystic fibrosis (CF) produce thick mucus that clogs airways and provides a very favorable environment for infection by bacteria that further decrease lung function and, ultimately, mortality. CF patients are often infected by bacteria such as Pseudomonas aeruginosa early in life and experience a series of chronic infections that, over time, become increasingly difficult to treat due to increased antibiotic resistance. Colistin is a major antibiotic used to treat CF patients. Clinical and laboratory studies have identified PmrA/PmrB and PhoP/PhoQ as responsible for increased resistance to colistin. Both have been identified in CF patient lungs, but why, in some cases, is it one and not the other? In this study, we show that distinct evolutionary trajectories to colistin resistance may be favored by the microaerobic partitioning found within the damaged CF lung.

Osmosis-induced hydrodynamic centering of W/O/W double emulsion droplets for quasi-concentric microcapsule/microsphere fabrication

Water-in-oil-in-water (W/O/W) double emulsion droplets are a typical colloidal system applied in various practical fields. However, certain high-specification applications necessitate exact geometric homogeneity in terms of both shell diameter and thickness. Despite achieving uniformity in shell diameter, on-chip microfluidic emulsification always leads to an eccentric phenomenon due to innate density differences of the droplet solution. At the same time, the current regulation methods have multiple limitations, including their applicability, regulatory scope, and objective restrictions. Herein, we propose an off-chip osmosis-induced hydrodynamic method to accurately regulate the eccentricity of W/O/W double emulsion droplets. The rational selection of hypertonic osmosis can energize the droplets and induce inward transmembrane water flux. This flow helps to reposition the droplet core to align concentrically with the shell, regardless of the initial dimension and eccentricity of the droplets. As the osmotic induction reduces due to the dilution of the inner solute molar concentration, the optimal droplet configuration can be achieved when osmosis-induced hydrodynamic regulation sufficiently counteracts the adverse effects of droplet solution density difference. The microcapsules can thus be produced by instant UV polymerization of the regulated double emulsion droplets, and the silica microspheres with fine geometric homogeneity can be prepared through further heat treatment, which is expected to meet the stringent morphological requirements in practical applications.

Establishment and Validation of an Integrated Microfluidic Step Emulsification Chip Supporting Droplet Digital Nucleic Acid Analysis.

Uniform and stable droplet generation is critical for accurate and efficient digital nucleic acid analysis (dNAA). In this study, an integrated microfluidic step emulsification device with wide-range droplet generation capability, small device dimensions, convenient fabrication strategy, low contamination and high robustness was developed. A tree-shaped droplet generation nozzle distribution design was proposed to increase the uniformity of droplet generation by equating flow rates, and the flow field in the design was numerically simulated. Theoretical analysis and comparative experiments on droplet size were performed regarding the influences of nozzle dimensions and surface properties. With incubation and hydrophobic reagent treatment, droplets as small as 73.1 μm were generated with multiplex nozzles of 18 μm (h) × 80 μm (w). The droplets were then collected into a standard PCR tube and an on-chip monolayer droplet collection chamber, without manual transfer and sample contamination. The oil-to-sample volume ratio in the PCR tube was recorded during collection. In the end, the droplets generated and collected using the microfluidic device proved to be stable and uniform for nucleic acid amplification and detection. This study provides reliable characteristic information for the design and fabrication of a micro-droplet generation device, and represents a promising approach for the realization of a three-in-one dNAA device under a step emulsification method.

Droplet-Based Microfluidics as a Platform to Design Food-Grade Delivery Systems Based on the Entrapped Compound Type.

Microfluidic technology has emerged as a powerful tool for several applications, including chemistry, physics, biology, and engineering. Due to the laminar regime, droplet-based microfluidics enable the development of diverse delivery systems based on food-grade emulsions, such as multiple emulsions, microgels, microcapsules, solid lipid microparticles, and giant liposomes. Additionally, by precisely manipulating fluids on the low-energy-demand micrometer scale, it becomes possible to control the size, shape, and dispersity of generated droplets, which makes microfluidic emulsification an excellent approach for tailoring delivery system properties based on the nature of the entrapped compounds. Thus, this review points out the most current advances in droplet-based microfluidic processes, which successfully use food-grade emulsions to develop simple and complex delivery systems. In this context, we summarized the principles of droplet-based microfluidics, introducing the most common microdevice geometries, the materials used in the manufacture, and the forces involved in the different droplet-generation processes into the microchannels. Subsequently, the encapsulated compound type, classified as lipophilic or hydrophilic functional compounds, was used as a starting point to present current advances in delivery systems using food-grade emulsions and their assembly using microfluidic technologies. Finally, we discuss the limitations and perspectives of scale-up in droplet-based microfluidic approaches, including the challenges that have limited the transition of microfluidic processes from the lab-scale to the industrial-scale.

High-Throughput Microfluidic Production of Droplets and Hydrogel Microspheres through Monolithically Integrated Microchannel Plates.

In this paper, we present a highly effective microfluidic emulsion system using an integrated microchannel plate (MCP), a porous glass membrane that is readily available and densely packs millions of through-microchannels, for high-throughput production of monodisperse droplets. The physical controls of droplet formation, including viscosity, flow rate, and pore size, have been extensively explored for optimum emulsification conditions. The performance of the device has been validated where monodisperse droplets with a narrow coefficient of variance (<5%) can be achieved at a dispersed phase flux of 3 mL h-1 from a piece of 4 × 4 mm2 MCP. The average droplet size is two times the nominal membrane pore diameter and thus can be easily controlled by choosing the appropriate membrane type. The preparation of hydrogel microspheres has also been demonstrated with a high throughput of 1.5 × 106 particles min-1. These microspheres with a uniform size range and rough surface morphology provide suitable bioenvironments and serve as ideal carriers for cell culture. Mouse fibroblasts are shown to be cultured on these 3D scaffolds with an average cell viability of over 96%. The cell attachment rate can reach up to 112 ± 7% in 24 h and the proliferation ability increases with the number of culture days. Furthermore, the device has been applied in the droplet digital polymerase chain reaction for absolute quantification of lung cancer-related PLAU genes. The detection limit achieved was noted to be 0.5 copies/μL with a dynamic range of 105 ranging from 1 × 102 to 1 × 106 copies/μL. Given the easy fabrication, robust performance, and simple operation, the emulsion system sets the stage for the laboratory's droplet-based assays and applications in tissue engineering.

Recent Progress of Droplet Microfluidic Emulsification Based Synthesis of Functional Microparticles.

The remarkable control function over the functional material formation process enabled by droplet microfluidic emulsification approaches can lead to the efficient and one-step encapsulation of active substances in microparticles, with the microparticle characteristics well regulated. In comparison to the conventional fabrication methods, droplet microfluidic technology can not only construct microparticles with various shapes, but also provide excellent templates, which enrich and expand the application fields of microparticles. For instance, intersection with disciplines in pharmacy, life sciences, and others, modifying the structure of microspheres and appending functional materials can be completed in the preparation of microparticles. The as-prepared polymer particles have great potential in a wide range of applications for chemical analysis, heavy metal adsorption, and detection. This review systematically introduces the devices and basic principles of particle preparation using droplet microfluidic technology and discusses the research of functional microparticle formation with high monodispersity, involving a plethora of types including spherical, nonspherical, and Janus type, as well as core-shell, hole-shell, and controllable multicompartment particles. Moreover, this review paper also exhibits a critical analysis of the current status and existing challenges, and outlook of the future development in the emerging fields has been discussed.

Robust generation of monodisperse droplets using a microfluidic step emulsification device with triangular nozzle

AbstractIn this work, the droplet generation process in the microfluidic step emulsification chip with a triangular nozzle (SE‐T) was investigated in the combination of visualization experiment and numerical simulation, through a comparison with a rectangular nozzle (SE‐R). The flow regimes, including dripping, dripping‐jetting transition, and jetting, were observed in the SE‐T, among which the dripping is the preferred flow regime to generate monodispersed droplet with corresponding C.V. (coefficient of variation) of the droplet size smaller than 1.9%. Compared with the SE‐R, the larger space and expanding structure of the triangular nozzle in the SE‐T enhance the wall wetting effects, which induces earlier appearance and accelerates shrinking of the neck. As a result, the SE‐T exhibits more robust droplet performance under the dripping regime, which produces the droplets with nearly unchanged size and higher monodispersity, especially little related to the variations of surfactant concentrations and dispersed phase flow rates.