Continuous Flow Microfluidic Device Research Articles

"On-The-Fly" Synthesis of Self-Supported LDH Hollow Structures Through Controlled Microfluidic Reaction-Diffusion Conditions.

Layered double hydroxides (LDHs) are a class of functional materials that exhibit exceptional properties for diverse applications in areas such as heterogeneous catalysis, energy storage and conversion, and bio-medical applications, among others. Efforts have been devoted to produce millimeter-scale LDH structures for direct integration into functional devices. However, the controlled synthesis of self-supported continuous LDH materials with hierarchical structuring up to the millimeter scale through a straightforward one-pot reaction method remains unaddressed. Herein, it is shown that millimeter-scale self-supported LDH structures can be produced by means of a continuous flow microfluidic device in a rapid and reproducible one-pot process. Additionally, the microfluidic approach not only allows for an "on-the-fly" formation of unprecedented LDH composite structures, but also for the seamless integration of millimeter-scale LDH structures into functional devices. This method holds the potential to unlock the integrability of these materials, maintaining their performance and functionality, while diverging from conventional techniques like pelletization and densification that often compromise these aspects. This strategy will enable exciting advancements in LDH performance and functionality.

Diffusive–layered convection in Y-shaped continuous-flow microreactor

Efficient and rapid mixing of fluids in continuous-flow microfluidic devices plays an important role in a wide range of applications. Although there are many different methods to enhance mixing in such reactors, the potential of natural convection to mix fluids has been overlooked because of its possible limitations at the microscale. Recent studies have challenged this statement by demonstrating the significance of both gravity-dependent and gravity-independent types of natural convection in improving mixing efficiency. It was shown that inducing double-diffusive convection, Marangoni convection, or Rayleigh–Taylor convection can greatly enhance the mixing of two aqueous solutions in a continuous-flow microreactor. In this study, we experimentally investigate the influence of another type of differential-diffusion instability, known as diffusive–layered convection, on the mixing of two aqueous solutions of chemically inert substances in a Y-shaped continuous-flow microreactor. We consider the scenario when two miscible solutions flow into the main channel through distinct tubes, in which they come into contact and mix, thus causing the onset of diffusive–layered convection. We used two experimental methods, including laser interferometry and dye visualization, to analyse the mixing process. Surprisingly, despite the vigorous convective motion driven by diffusive–layered convection, the intensity of mixing of pumped fluids remains comparable to the scenario when solutions mix only via a diffusion mechanism. We explain this unexpected result by the unique structure of the convective motion, which prevents system homogenization. A comparison with the results previously obtained for double-diffusive convection is performed, and pure diffusive mixing scenarios are implemented.

A di-electrophoretic simulation procedure of iron-oxide micro-particle drug attachment system for leukemia treatment using COMSOL software: a potential treatment reference for LMICs.

Leukemia encompasses various subtypes, each with unique characteristics and treatment approaches. The challenge lies in developing targeted therapies that can effectively address the specific genetic mutations or abnormalities associated with each subtype. Some leukemia cases may become resistant to existing treatments over time making them less susceptible to chemotherapy or other standard therapies. Developing new treatment strategies to overcome resistance is an ongoing challenge particularly in Low and Middle Income Countries (LMICs). Computational studies using COMSOL software could provide an economical, fast and resourceful approach to the treatment of complicated cancers like leukemia. Using COMSOL Multiphysics software, a continuous flow microfluidic device capable of delivering anti-leukemia drugs to early-stage leukemia cells has been computationally modeled using dielectrophoresis (DEP). The cell size difference enabled the micro-particle drug attachment to the leukemia cells using hydrodynamic focusing from the dielectrophoretic force. This point of care application produced a low voltage from numerically calculated electrical field and flow speed simulations. Therefore, such a dielectrophoretic low voltage application model can be used as a computational treatment reference for early-stage leukemia cells with an approximate size of 5 μm.

Innenrücktitelbild: Highly Efficient Self‐Assembly of Metallacages and Their Supramolecular Catalysis Behaviors in Microdroplets (Angew. Chem. 27/2023)

A highly efficient strategy for achieving the supramolecular self-assembly of well-defined metallacages in microdroplets through continuous-flow microfluidic devices is demonstrated by Lin Xu et al. in their Research Article (e202301900). The strategy adds a highly efficient approach to the toolbox of metallacage self-assembly.

Inside Back Cover: Highly Efficient Self‐Assembly of Metallacages and Their Supramolecular Catalysis Behaviors in Microdroplets (Angew. Chem. Int. Ed. 27/2023)

A highly efficient strategy for achieving the supramolecular self-assembly of well-defined metallacages in microdroplets through continuous-flow microfluidic devices is demonstrated by Lin Xu et al. in their Research Article (e202301900). The strategy adds a highly efficient approach to the toolbox of metallacage self-assembly.

Highly Efficient Self-Assembly of Metallacages and Their Supramolecular Catalysis Behaviors in Microdroplets.

Developing a new strategy to improve the self-assembly efficiency of functional assemblies in a confined space and construct hybrid functional materials is a significant and fascinating endeavor. Herein, we present a highly efficient strategy for achieving the supramolecular self-assembly of well-defined metallacages in microdroplets through continuous-flow microfluidic devices. The high efficiency and versatility of this approach are demonstrated by the generation of five representative metallacages in different solvents containing water, DMF, acetonitrile, and methanol in a few minutes with nearly quantitative yields, in contrast to the yields obtained with the hour-scale reaction time in a batch reactor. A ring-opening catalytic reaction of the metallacages was selected as a model reaction for exploring supramolecular catalysis in microdroplets, whereby the catalytic yield was enhanced by 2.22-fold compared to that of the same reaction in the batch reactor. This work illustrates a new promising approach for the self-assembly of supramolecular systems.

A Method for Rapid, Quantitative Evaluation of Particle Sorting in Microfluidics Using Basic Cytometry Equipment

This paper describes, in detail, a method that uses flow cytometry to quantitatively characterise the performance of continuous-flow microfluidic devices designed to separate particles. Whilst simple, this approach overcomes many of the issues with the current commonly utilised methods (high-speed fluorescent imaging, or cell counting via either a hemocytometer or a cell counter), as it can accurately assess device performance even in complex, high concentration mixtures in a way that was previously not possible. Uniquely, this approach takes advantage of pulse processing in flow cytometry to allow quantitation of cell separation efficiencies and resulting sample purities on both single cells as well as cell clusters (such as circulating tumour cell (CTC) clusters). Furthermore, it can readily be combined with cell surface phenotyping to measure separation efficiencies and purities in complex cell mixtures. This method will facilitate the rapid development of a raft of continuous flow microfluidic devices, will be helpful in testing novel separation devices for biologically relevant clusters of cells such as CTC clusters, and will provide a quantitative assessment of device performance in complex samples, which was previously impossible.

Percolation-assisted coating of metal-organic frameworks on porous substrates

In the past two decades, significant advancements have been made toward thin film fabrication of metal-organic frameworks (MOFs) on porous solid substrates for gas separation and catalysis applications. To enhance the physical and chemical stability of the films, various top-down and bottom-up approaches have been implemented. While both approaches have some advantages, they are mostly limited due to a lack of adhesion with the substrate, cracking of the films, and non-uniform coverages. Therefore, there is a need for improvement in the coating processes for the fabrication of robust, uniform, and scalable films. Here, we develop a novel percolation-assisted coating (PAC) process that combines top-down and bottom-up approaches in a continuous-flow microfluidic device to deposit HKUST-1 on a porous substrate, yielding controlled film thicknesses and mass loading. The PAC process is optimized using a Multiphysics modeling approach to design the microfluidic reactor, which can be readily scaled up and deployed to fabricate MOF films on various porous substrates. The desired thickness in the range of 10–200 μm of the MOF film can be achieved by controlling the residence time and temperature of the reaction mixture. The synthesized film is characterized for physical adhesion using sonication, film coverage using scanning electron microscopy, and porosity using a porosimeter. The performance of synthesized films is benchmarked for the effective separation of 50 vol% CH4–H2 gas mixture with the separation factor of 4. The microfluidic device is also applicable to synthesize films of various MOFs like MOF-5, MOF-505 (also reported here), UIO-66, etc., over a wide range of porous substrates. Lastly, we propose a multi-chamber design of the microfluidic device for high-throughput screening of thin-film growth using the PAC process.

Covalent transfer of chemical gradients onto a graphenic surface with 2D and 3D control

Control over the functionalization of graphenic materials is key to enable their full application in electronic and optical technologies. Covalent functionalization strategies have been proposed as an approach to tailor the interfaces’ structure and properties. However, to date, none of the proposed methods allow for a covalent functionalization with control over the grafting density, layer thickness and/or morphology, which are key aspects for fine-tuning the processability and performance of graphenic materials. Here, we show that the no-slip boundary condition at the walls of a continuous flow microfluidic device offers a way to generate controlled chemical gradients onto a graphenic material with 2D and 3D control, a possibility that will allow the sophisticated functionalization of these technologically-relevant materials.

Recent approaches to mRNA vaccine delivery by lipid-based vectors prepared by continuous-flow microfluidic devices.

Advancements in nanotechnology have resulted in the introduction of several nonviral delivery vectors for the nontoxic, efficient delivery of encapsulated mRNA-based vaccines. Lipid- and polymer-based nanoparticles (NP) have proven to be the most potent delivery systems, providing increased delivery efficiency and protection of mRNA molecules from degradation. Here, the authors provide an overview of the recent studies carried out using lipid NPs and their functionalized forms, polymeric and lipid-polymer hybrid nanocarriers utilized mainly for the encapsulation of mRNAs for gene and immune therapeutic applications. A microfluidic system as a prevalent methodology for the preparation of NPs with continuous flow enables NP size tuning, rapid mixing and production reproducibility. Continuous-flow microfluidic devices for lipid and polymeric encapsulated RNA NP production are specifically reviewed.

On the efficiency of convective mixing in a Y-shaped channel

Continuous-flow microfluidic devices are widely used in microbiology, fine organic synthesis, pharmaceuticals, biomedicine, etc. Most applications require rapid mixing of the fluids that pass through the microfluidic chip. The mechanism of natural diffusion is not always efficient due to limitations on the length of the channel. In this work, we numerically study the efficiency of using various mechanisms of natural convection for the mixing of fluids entering the microfluidic chip. Solutions typically differ in buoyancy and diffusion rates of dissolved components, making them sensitive to gravity-dependent instabilities such as Rayleigh-Taylor convection, double diffusion and diffusion layer convection. We consider a Y-shaped microchannel, which is, on the one hand, the simplest, and, on the other hand, a typical element of a microfluidic network. We assume that two miscible solutions independently flow into a common channel where they come into contact. For each type of instability, we numerically estimated the characteristic channel length, after which complete mixing of the solutions occurs. The simulations were performed in the framework of both 2D and 3D models. Finally, we compare the numerical results with the experimental data obtained recently.

Environmentally Friendly Method of Assembly of Cardanol and Cholesterol into Nanostructures Using a Continuous Flow Microfluidic Device

This study shows a viable and straightforward microfluidic method of assembly of cardanol (CA) and cholesterol (CH) into amphiphile nanostructures obtained through a hydrodynamic focusing approach according to which an alcoholic solution of CA and CH is constrained within a two-dimensional lamina shape by two lateral streams of borate buffer solution. The process is performed within glass-made cross-shaped micro-sized fluidic chips specially designed to achieve a laminar regime. CA, distilled from the cashew nut shell liquid, is demonstrated as a surface-active molecule in borate buffer basic medium and when mixed with CH it produces versatile nanovesicles through an in-batch solvent-free process. Compared to this conventional method, the microfluidic route allows operating under continuous flows, with a reduced amount of reagents and at lower experimental temperatures, ensuring no waste formation and the achievement of size-monodisperse amphiphile nanostructures that do not need further steps of purification. Electron microscopy analyses demonstrate that upon increasing CH in the lipid mixture, a switchover from spherical CA micelles to CA/CH mixed closed vesicles occurs. Differential scanning microcalorimetry confirms the formation of vesicular structures and evidences the primary role of CH, which increasingly lowers the temperature of transition depending on its concentration.

Trojan pH-Sensitive Polymer Particles Produced in a Continuous-Flow Capillary Microfluidic Device Using Water-in-Oil-in-Water Double-Emulsion Droplets.

A facile and robust microfluidic method to produce nanoparticle-in-microparticle systems (Trojan systems) is reported as a delivery vector for the oral administration of active pharmaceutical ingredients. The microfluidic system is based on two coaxial capillaries that produce monodisperse water-in-oil-in-water (W/O/W) double emulsions in a highly controlled fashion with precise control over the resulting particle structure, including the core and shell dimensions. The influence of the three phase flow rates, pH and drying process on the formation and overall size is evaluated. These droplets are then used as templates for the production of pH-sensitive Trojan microparticles after solvent evaporation. The shell of Trojan microparticles is made of Eudragit®, a methacrylic acid-ethyl acrylate copolymer that would enable the Trojan microparticle payload to first pass through the stomach without being degraded and then dissolve in the intestinal fluid, releasing the inner payload. The synthesis of the pH-sensitive Trojan microparticles was also compared with a conventional batch production method. The payloads considered in this work were different in nature: (1) fluorescein, to validate the feasibility of the polymeric shell to protect the payload under gastric pH; (2) poly(D,L-lactic acid/glycolic acid)-PLGA nanoparticles loaded with the antibiotic rifampicin. These PLGA nanoparticles were produced also using a microfluidic continuous process and (3) PLGA nanoparticles loaded with Au nanoparticles to trace the PLGA formulation under different environments (gastric and intestinal), and to assess whether active pharmaceutical ingredient (API) encapsulation in PLGA is due efficiently. We further showed that Trojan microparticles released the embedded PLGA nanoparticles in contact with suitable media, as confirmed by electron microscopy. Finally, the results show the possibility of developing Trojan microparticles in a continuous manner with the ability to deliver therapeutic nanoparticles in the gastrointestinal tract.

On-the-spot quenching for effective implementation of cooling crystallization in a continuous-flow microfluidic device

Illustrated is a microfludic cooling crystallization device that can effectively screen polymorphs, growth rates, and morphology of crystalline materials.

In-line measurement of liquid-liquid phase separation boundaries using a turbidity-sensor-integrated continuous-flow microfluidic device.

Liquid-liquid phase separation (LLPS), also known as oiling-out, is the appearance of the second liquid phase preceding the crystallization. LLPS is an undesirable phenomenon that can occur during the crystallization of active pharmaceutical ingredients (APIs), proteins, and polymers. It is typically avoided during crystallization due to its detrimental impacts on crystalline products due to lowered crystallization rate, the inclusion of impurities, and alteration in particle morphology and size distribution. In situ monitoring of phase separation enables investigating LLPS and identifying the phase separation boundaries. Various process analytical technologies (PATs) have been implemented to determine the LLPS boundaries prior to crystallization to prevent oiling out of compounds. The LLPS measurements using PATs can be time-consuming, expensive, and challenging. Here, we have implemented a fully integrated continuous-flow microfluidic device with a turbidity sensor to quickly and accurately evaluate the LLPS boundaries for a β-alanine, water, and IPA mixture. The turbidity-sensor-integrated continuous-flow microfluidic device is also placed under an optical microscope to visually track and record the appearance and disappearance of oil droplets. Streams of an aqueous solution of β-alanine, pure solvent (water), and pure antisolvent (IPA or ethanol) are pumped into the continuous-flow microfluidic device at various flow rates to obtain the compositions at which the solution becomes turbid. The onset of turbidity is measured using a custom-designed, in-line turbidity sensor. The LLPS boundaries can be estimated using the turbidity-sensor-integrated microfluidic device in less than 30 min, which will significantly improve and enhance the workflow of the pharmaceutical drug (or crystalline material) development process.

A circular gradient-width crossflow microfluidic platform for high-efficiency blood plasma separation

Separation and collection of blood plasma are required steps for broad ranges of clinical diagnostic assays. Here, we introduce a continuous-flow blood plasma separation microfluidic device based on size exclusion cell separation in crossflow microchannels. A gradient-width crossflow microchannel structure is utilized to minimize blood cell leakage into the plasma collection channel while maximizing the flow rate for higher throughput. Additionally, the crossflow channels are placed at the corner of the blood channel to enhance Fahraeus effects, resulting in reduction of the concentration of blood cells around the entrance of the crossflow channels. Combined with a circular geometry that minimizes the footprint of the device for point-of-care diagnosis applications, the developed device significantly improves the performance of blood plasma separation in terms of separation efficiency, degree of hemolysis, and microchannel clogging by blood cells. At a relatively high flow rate of 2000 µl/h for a passive blood separation device, a plasma extraction ratio of 22% and red blood cell separation efficiency of 97% could be achieved using undiluted whole human blood, where such separation efficiency is about 28% higher compared with that of a conventional crossflow design. The effect of channel height and flow rate on the degree of hemolysis was also investigated, where the optimized device could achieve very little hemolysis (0.01–0.5%), demonstrating that the device can produce high-quality blood plasma. Integration of this high-performance, compact, and passive blood plasma separation microdevice with on-chip blood biomarker sensing elements can create a point-of-care diagnosis system with reduced cost, test time, and sample/reagent volume.

A straight-forward, inexpensive, low power continuous-flow µPCR chip using PCB-based heater electrodes with uniform temperature distribution

A relatively high-efficiency, two-step type continuous flow microfluidic device for polymerase chain reaction (PCR) is presented in this work. The heater electrodes are designed and fabricated based on 0.25 µm-thick two-layer standard printed circuit board (PCB) technology. Temperature uniformity in different temperature zones of PCR plays a key role in increasing process efficiency. By properly designing meander shaped electrodes, relatively good temperature uniformity and high PCR efficiency is realized. A symmetrical structure is demonstrated for the arrangement of the electrodes, where the annealing electrode is placed between two denaturation electrodes. In this way, we have favorably used the thermal cross talk between the two temperature zones in order to create a relatively uniform temperature in the annealing electrode. Heat spreaders between meander lines in denaturation electrodes are used to distribute temperature uniformly, in contrast to the literature, where complex optimization methods are adopted. This eliminates the need to use bulky heat spreaders such as copper or aluminum blocks which are commonly used to uniformize temperature in continuous flow µPCRs. As a result, the number of components and the overall size of the device and consequently the complexity and fabrication cost decreases. On the other hand, the thermal mass is reduced, which in turn reduces the power consumption of the system. The temperature measurements are first performed with contact PT100 temperature sensors and then the results are verified by a thermal camera. The temperature difference measured by PT100 sensors approximately matches the standard deviation obtained from the thermal camera in a specific area. For the denaturation electrode these values are 1.14 °C and 1.2 °C, respectively. A maximum absolute temperature difference of 1 °C is measured for the annealing electrode. The fabricated microchip with dimensions of 50 mm × 40 mm is tested by an 88 base-pair DNA sample from the human housekeeping gene with a power consumption of less than 0.82 W. A relatively remarkable amplification efficiency is achieved to prove the efficacy of our inexpensive PCB electrodes.

Miniaturized DNA amplification platform with soft-lithographically fabricated continuous-flow PCR microfluidic device on a portable temperature controller

Miniaturized DNA amplification platform with soft-lithographically fabricated continuous-flow PCR microfluidic device on a portable temperature controller

Exploiting Reaction-Diffusion Conditions to Trigger Pathway Complexity in the Growth of a MOF.

Coordination polymers (CPs), including metal-organic frameworks (MOFs), are crystalline materials with promising applications in electronics, magnetism, catalysis, and gas storage/separation. However, the mechanisms and pathways underlying their formation remain largely undisclosed. Herein, we demonstrate that diffusion-controlled mixing of reagents at the very early stages of the crystallization process (i.e., within ≈40 ms), achieved by using continuous-flow microfluidic devices, can be used to enable novel crystallization pathways of a prototypical spin-crossover MOF towards its thermodynamic product. In particular, two distinct and unprecedented nucleation-growth pathways were experimentally observed when crystallization was triggered under microfluidic mixing. Full-atom molecular dynamics simulations also confirm the occurrence of these two distinct pathways during crystal growth. In sharp contrast, a crystallization by particle attachment was observed under bulk (turbulent) mixing. These unprecedented results provide a sound basis for understanding the growth of CPs and open up new avenues for the engineering of porous materials by using out-of-equilibrium conditions.

Inside Cover: Exploiting Reaction‐Diffusion Conditions to Trigger Pathway Complexity in the Growth of a MOF (Angew. Chem. Int. Ed. 29/2021)

How do you unveil pathway complexity in a crystallization process? In their Research Article on page 15920, Alessandro Sorrenti, Marco D′Abramo, Guillermo Mínguez Espallargas, Josep Puigmartí-Luis, and co-workers show that harnessing a reaction–diffusion (RD) process within a continuous flow microfluidic device, and on a millisecond timescale, is key to enable two unprecedented nucleation–growth pathways during a MOF synthesis.