Microfluidic Design Research Articles

Innovative, Flexible, and Miniaturized Microfluidic Paper-Based Plasmonic Chip for Efficient Near-Infrared Metal Enhanced Fluorescence Biosensing and Imaging.

The implementation of metal enhanced fluorescence (MEF) as an efficient detection tool, especially in the near-infrared region of the electromagnetic spectrum, is a rather new direction for diagnostic analytical technologies. In this context, we propose a novel microfluidic plasmonic design based on paper for efficient MEF detection of the "proof-of-concept" biotin-streptavidin recognition interaction. Our design made use of the benefits of gold nanobipyramids (AuBPs), considering the strong enhanced electromagnetic field present at their sharp tips, and filter paper to operate as a natural microfluidic channel due to excellent wicking abilities. The calligraphed plasmonic paper, obtained using a commercial pen filled with AuBPs, was integrated in a robust sandwich optically transparent polydimethylsiloxane chip, exhibiting portability and flexibility while preserving the chip's properties. To place the Alexa 680 fluorophore at an optimal distance from the nanobipyramid substrate, the human IgG-anti-IgG-conjugated biotin sandwich reaction was employed. Thus, upon the capture of Alexa 680-conjugated streptavidin by the biotinylated system, a 1.3-fold average enhancement of the fluorophore's emission was determined by bulk fluorescence measurements. However, the local enhancement factor was considerably higher with values spanning from 5 to 6.3, as proven by mapping the fluorescence emission under both re-scan microscopy and fluorescence lifetime imaging, endorsing the proposed chip's feasibility for bulk MEF biosensing as well as high-resolution MEF bioimaging. Finally, the versatility of our chip was demonstrated by adapting the biosensing protocol for cardiac troponin I biomarker detection, validated using 10 plasma samples collected from pediatric patients and corroborated with a conventional ELISA assay.

Wettability effect on displacement in disordered media under preferential flow conditions

Preferential flow in a porous medium is commonly encountered in many practical applications. Our previous studies have discovered the preferential flow-induced non-monotonic wettability effect on displacement (J. Fluid Mech, vol. 942, 2022a, R5), but whether this non-monotonic rule is consistent for different disordered media and the impact of the interplay between the disorder and wettability under preferential flow conditions is still not well understood. Here, we combine microfluidic experiments, pore-scale simulations and theoretical analysis to study the impact of the disorder on the invading process where wettability varies from strongly water wet to strongly oil wet. Even though the strongly disordered matrix varies to a uniform state, the generality of the preferential flow-induced non-monotonic wettability effect is still proved. However, the previous pore-scale dynamics based on the strongly disordered matrix cannot explain the invading behaviour in the uniform matrix under preferential flow conditions. New mechanisms for the uniform matrix are further investigated by pore-scale modelling, which indicates that the balance of microscopic imbibition ability and the macroscopic interfacial stability dominate the invading process. We derive a theoretical model to describe the wettability effect and predict the optimal contact angle, which fits well with experimental and simulation results. Our work extends the understanding of the impact of preferential flow conditions on the wettability effect and is also of practical significance for engineering applications, such as geological CO2 sequestration, enhanced hydrocarbon recovery, soil wetting, liquid-infused material fabrication and microfluidic device design.

Ultra-Fast Ion Mobility Spectrometer for High-Throughput Chromatography.

Fast chromatography systems especially developed for high sample throughput applications require sensitive detectors with a high repetition rate. These high throughput techniques, including various chip-based microfluidic designs, often benefit from detectors providing subsequent separation in another dimension, such as mass spectrometry or ion mobility spectrometry (IMS), giving additional information about the analytes or monitoring reaction kinetics. However, subsequent separation is required at a high repetition rate. Here, we therefore present an ultra-fast drift tube IMS operating at ambient pressure. Short drift times while maintaining high resolving power are reached by several key instrumental design features: short length of the drift tube, resistor network of the drift tube, tristate ion shutter, and improved data acquisition electronics. With these design improvements, even slow ions with a reduced mobility of just 0.94 cm2/(V s) have a drift time below 1.6 ms. Such short drift times allow for a significantly increased repetition rate of 600 Hz compared with previously reported values. To further reduce drift times and thus increase the repetition rate, helium can be used as the drift gas, which allows repetition rates of up to 2 kHz. Finally, these significant improvements enable IMS to be used as a detector following ultra-fast separation including chip-based chromatographic systems or droplet microfluidic applications requiring high repetition rates.

Capturing the Dynamic Processes of Porosity Clogging

AbstractUnderstanding mineral precipitation induced porosity clogging and being able to quantify its non‐linear feedback on transport properties is fundamental for predicting the long‐term evolution of energy‐related subsurface systems. Commonly applied porosity‐diffusivity relations used in numerical simulations on the continuum‐scale predict the case of clogging as a final state. However, recent experiments and pore‐scale modeling investigations suggest dissolution‐recrystallization processes causing a non‐negligible inherent diffusivity of newly formed precipitates. To verify these processes, we present a novel microfluidic reactor design that combines time‐lapse optical microscopy and confocal Raman spectroscopy, providing real‐time insights of mineral precipitation induced porosity clogging under purely diffusive transport conditions. Based on 2D optical images, the effective diffusivity was determined as a function of the evolving porous media, using pore‐scale modeling. At the clogged state, Raman isotopic tracer experiments were conducted to visualize the transport of deuterium through the evolving microporosity of the precipitates, demonstrating the non‐final state of clogging. The evolution of the porosity‐diffusivity relationship in response to precipitation reactions shows a behavior deviating from Archie's law. The application of an extended power law improved the description of the evolving porosity‐diffusivity, but still neglected post‐clogging features. Our innovative combination of microfluidic experiments and pore‐scale modeling opens new possibilities to validate and identify relevant pore‐scale processes, providing data for upscaling approaches to derive key relationships for continuum‐scale reactive transport simulations.

Molecular Separation by Using Active and Passive Microfluidic chip Designs: A Comprehensive Review

AbstractSeparation and identification of molecules and biomolecules such as nucleic acids, proteins, and polysaccharides from complex fluids are known to be important due to unmet needs in various applications. Generally, many different separation techniques, including chromatography, electrophoresis, and magnetophoresis, have been developed to identify the target molecules precisely. However, these techniques are expensive and time consuming. “Lab‐on‐a‐chip” systems with low cost per device, quick analysis capabilities, and minimal sample consumption seem to be ideal candidates for separating particles, cells, blood samples, and molecules. From this perspective, different microfluidic‐based techniques have been extensively developed in the past two decades to separate samples with different origins. In this review, “lab‐on‐a‐chip” methods by passive, active, and hybrid approaches for the separation of biomolecules developed in the past decade are comprehensively discussed. Due to the wide variety in the field, it will be impossible to cover every facet of the subject. Therefore, this review paper covers passive and active methods generally used for biomolecule separation. Then, an investigation of the combined sophisticated methods is highlighted. The spotlight also will be shined on the elegance of separation successes in recent years, and the remainder of the article explores how these permit the development of novel techniques.

Analytical Validation of a Spiral Microfluidic Chip with Hydrofoil-Shaped Pillars for the Enrichment of Circulating Tumor Cells.

The isolation of circulating tumor cells (CTCs) from peripheral blood with high efficiency remains a challenge hindering the utilization of CTC enrichment methods in clinical practice. Here, we propose a microfluidic channel design for the size-based hydrodynamic enrichment of CTCs from blood in an epitope-independent and high-throughput manner. The microfluidic channel comprises a spiral-shaped part followed by a widening part, incorporating successive streamlined pillars, that improves the enrichment efficiency. The design was tested against two benchmark designs, a spiral microfluidic channel and a spiral microfluidic channel followed by a widening channel without the hydrofoils, by processing 5 mL of healthy blood samples spiked with 100 MCF-7 cells. The results proved that the design with hydrofoil-shaped pillars perform significantly better in terms of recovery (recovery rate of 67.9% compared to 23.6% in spiral and 56.7% in spiral with widening section), at a cost of slightly lower white blood cell (WBC) depletion (depletion rate of 94.2% compared to 98.6% in spiral and 94.2% in spiral with widening section), at 1500 µL/min flow rate. For analytical validation, the design was further tested with A549, SKOV-3, and BT-474 cell lines, yielding recovery rates of 62.3 ± 8.4%, 71.0 ± 6.5%, and 82.9 ± 9.9%, respectively. The results are consistent with the size and deformability variation in the respective cell lines, where the increasing size and decreasing deformability affect the recovery rate in a positive manner. The analysis before and after the microfluidic chip process showed that the process does not affect cell viability.

One Step Encapsulation of Mesenchymal Stromal Cells in PEG Norbornene Microgels for Therapeutic Actions.

Cell therapies require control over the cellular response under standardized conditions to ensure continuous delivery of therapeutic agents. Cell encapsulation in biomaterials can be particularly effective at providing cells with a uniformly supportive and permissive cell microenvironment. In this study, two microfluidic droplet device designs were used to successfully encapsulate equine mesenchymal stromal cells (MSCs) into photopolymerized polyethylene glycol norbornene (PEGNB) microscale (∼100-200 μm) hydrogel particles (microgels) in a single on-chip step. To overcome the slow cross-linking kinetics of thiol-ene reactions, long dithiol linkers were used in combination with a polymerization chamber customized to achieve precise retention time for microgels while maintaining cytocompatibility. Thus, homogeneous cell-laden microgels could be continuously fabricated in a high-throughput fashion. Varying linker length mediated both the gel formation rate and material physical properties (stiffness, mass transport, and mesh size) of fabricated microgels. Postencapsulation cell viability and therapeutic indicators of MSCs were evaluated over 14 days, during which the viability remained at least 90%. Gene expression of selected cytokines was not adversely affected by microencapsulation compared to monolayer MSCs. Notably, PEGNB-3.5k microgels rendered significant elevation in FGF-2 and TGF-β on the transcription level, and conditioned media collected from these cultures showed robust promotion in the migration and proliferation of fibroblasts. Collectively, standardized MSC on-chip encapsulation will lead to informed and precise translation to clinical studies, ultimately advancing a variety of tissue engineering and regenerative medicine practices.

Microfluidic Design of Streamlined Alginate Hydrogel Micromotors with Run and Tumble Motion Patterns.

Autonomous micromotors demonstrate remarkable advancements in biomedical applications. A noteworthy example is streamlined motors, which display enhanced movement efficiency with low fluid-resistance. However, existing streamlined motors, primarily constructed from inorganic materials, present challenges due to their complex fabrication procedures and lack of a soft interface for interaction with biological systems. Herein, a novel design of biodegradable streamlined alginate hydrogel micromotors with a teardrop shape by microfluidics is introduced. The platform enables the high-throughput fabrication of monodisperse micromotors with varied dimensions. By incorporating Pt-coated Fe3 O4 nanoparticles, micromotors are equipped with dual capabilities of catalytic propulsion and accurate magnetic guidance. Through precisely tuning the localization regions of catalysts within the micromotors, the streamlined hydrogel micromotors not only exhibit enhanced propelling efficiency, but also accomplish distinct motion patterns of run and tumble. The design provides insights for developing advanced micromotors capable of executing intricate tasks across diverse application scenarios.

A 3D homogeneous microreactor with high mixing intensity at wide Re range for MOF preparation and POCT application

Most microreactors can only work under high Re or extremely low Re conditions to obtain enough mixing efficiency, the inefficient reaction intensity undoubtedly limits its application scenarios. In this study, a novel 3D homogeneous microreactor (3D-HM) was reported that demonstrate a strong mixing intensity over a wide range of Re conditions. The 3D geometries of shunt and lateral secondary flow structure were optimized in details, and then were added to the Zigzag mixing channel to promote mixing index (MI) (over 94 % from Re = 0–10, and more than 98 % for Re > 10). Compared with the traditional T-shaped, Split and Recombine micromixer, a shorter mixing distance (Lmix) and mixing time (τmix) were needed to realize fully mixing. The 3D-HM was used for the preparation of Zeolitic imidazolate framework (ZIF-8), the particle size, yield and drug loading capacity of ZIF-8 prepared using the 3D-HM under a milder Re conditions were better than those prepared by traditional and existing microfluidic methods under a harsh Re conditions. The promotion to diffusion mixing with 3D-HM channel was then verified using the enzymatic catalytic reaction driven by syringe pump at low Re and capillary pump, respectively. Enzyme and substrates in the laminar flow in the 3D-HM are more likely to bind each other than that in the T-shaped diffusion mixing. In addition, the laminar diffusion mixing process was scaled up by microfluidic channel design, so that the results of enzymic catalytic reaction was visualized for Point-of-care Testing (POCT) application. The MI, Lmix and τmix directly determine the homogeneous microreaction intensity. The proposal of the 3D-HM may provide more possibilities for the application of micromixing-based reactors.

A New Paper-Based Microfluidic Design (µPAD) for Determination of Total Phenols and Biological Applications Using Smartphone Sensor as a Detector

A New Paper-Based Microfluidic Design (µPAD) for Determination of Total Phenols and Biological Applications Using Smartphone Sensor as a Detector

Neural-physics multi-fidelity model with active learning and uncertainty quantification for GPU-enabled microfluidic concentration gradient generator design

The microfluidic concentration gradient generator (μCGG) is an important biomedical device to generate concentration gradients (CGs) of biomolecules at the microscale. Nonetheless, determining their operational parameter values to generate complex, user-specific, biologically desired CGs is not trivial. This paper presents a neural-physics multi-fidelity model (NP-MFM) to predict CGs with equivalent accuracy as high-fidelity CFD simulation at ultra-fast computational speed through a novel uncertainty- and distance-based active learning process. The verified NP-MFM, along with the genetic algorithm, is implemented on a GPU platform to search optimal values of operational parameters that generate CGs closely matching user-prescribed profiles. Results show that the NP-MFM is a feasible multi-fidelity modeling approach for rapid and accurate prediction of CGs (with 0.019s/simulation) and can be used for GPU-enabled μCGG design optimization and automation. Furthermore, design CGs generated by the proposed method match user-prescribed CGs very well with an averaged discrepancy less than 0.34.

Microfluidic design for in-vitro liver zonation-a numerical analysis using COMSOL Multiphysics.

The liver is one of the most important organs, with a complex physiology. Current in-vitro approaches are not accurate for disease modeling and drug toxicity research. One of those features is liver zonation, where cells display different physiological states due to different levels of oxygen and nutrient supplements. Organ-on-a-chip technology employs microfluidic platforms that enable a controlled environment for in-vitro cell culture. In this study, we propose a microfluidic design embedding a gas channel (of ambient air), creating an oxygen gradient. We numerically simulate different flow rates and cell densities with the COMSOL Multiphysics package considering cell-specific consumption rates of oxygen and glucose. We establish the cell density and flow rate for optimum oxygen and glucose distribution in the cell culture chamber. Furthermore, we show that a physiologically relevant concentration of oxygen and glucose in the chip is reached after 24h and 30min, respectively. The proposed microfluidic design and optimal parameters we identify in this paper provide a tool for in-vitro liver zonation studies. However, the microfluidic design is not exclusively for liver cell experiments but is foreseen to be applicable in cell studies where different gas concentration gradients are critical, e.g., studying hypoxia or toxic gas impact.

Stretchable Enzymatic Biofuel Cells Based on Microfluidic Structured Elastomeric Polydimethylsiloxane with Wrinkled Gold Electrodes

AbstractVarious sensors and electronic devices are recently developed to monitor human health in mechanically flexible or even stretchable forms for intimate contact with non‐flat curvilinear surfaces of the human body. For successful operation of these devices, finding a proper way to electrically power them is very important. In this work, glucose/oxygen fueled enzymatic biofuel cells (EBFCs) based on microfluidic structured elastomeric polydimethylsiloxane substrate with wrinkled gold (Au) electrodes are suggested for power supply. For doing that, firstly, bottom surface of microfluidic channel is covered with buckled Au electrodes for stretchability. By microfluidic design showing capillary imbibition through fluidic channels, loading of catalyts is promoted. Interestingly, buckled Au electrodes induce much better anodic and cathodic reaction rates than those of non‐buckled Au electrodes by 25% and 33%, respectively. This is because surface area and the amount of catalyst loading in electrodes increase by Au wrinkling. In evaluations of EBFCs using the buckled Au electrodes, maximum power density reaches 7.1 ± 0.64 µW cm−2, while they show decent performance of 5.4 ± 0.49 µW cm−2 even under external stretching. Taken together, it is corroborated that such proposed stretchable EBFCs are alternative for providing electrical power in wearable or implantable devices.

Freezing and thawing of cells on a microfluidic device: a simple and time-saving experimental procedure.

Developing cell cryopreservation methods on chips is not only crucial for biomedical science but also represents an innovative approach for preserving traditional cell samples. This study presents a simple method for direct cell freezing and thawing on chip, allowing for long-term storage of cells. During the freezing process, cells were injected into the microchannel along with a conventional cell cryopreservation solution, and the chip was packed using a self-sealing bag containing isopropyl alcohol and then stored in a -80°C refrigerator until needed. During the thawing process, microcolumn arrays with a spacing of 8µm were strategically incorporated into the microfluidic chip design to effectively inhibit cells from the channel. The breast cancer cell lines MDA-MB-231 and B47D demonstrated successful thawing and growth after cryopreservation for 1 month to 1 year. These findings offer a direct cell freezing and thawing method on a microfluidic chip for subsequent experiments.

Structural Optimization Design of Microfluidic Chips Based on Fast Sequence Pair Algorithm.

The market for microfluidic chips is experiencing significant growth; however, their development is hindered by a complex design process and low efficiency. Enhancing microfluidic chips' design quality and efficiency has emerged as an integral approach to foster their advancement. Currently, the existing structural design schemes lack careful consideration regarding the impact of chip area, microchannel length, and the number of intersections on chip design. This inadequacy leads to redundant chip structures resulting from the separation of layout and wiring design. This study proposes a structural optimization method for microfluidic chips to address these issues utilizing a simulated annealing algorithm. The simulated annealing algorithm generates an initial solution in advance using the fast sequence pair algorithm. Subsequently, an improved simulated annealing algorithm is employed to obtain the optimal solution for the device layout. During the wiring stage, an advanced wiring method is used to designate the high wiring area, thereby increasing the success rate of microfluidic chip wiring. Furthermore, the connection between layout and routing is reinforced through an improved layout adjustment method, which reduces the length of microchannels and the number of intersections. Finally, the effectiveness of the structural optimization approach is validated through six sets of test cases, successfully achieving the objective of enhancing the design quality of microfluidic chips.

Fluid dynamic design for mitigating undesired cell effects and its application to testis cell response testing to endocrine disruptors

Microfluidic devices have emerged as powerful tools for cell-based experiments, offering a controlled microenvironment that mimic the conditions within the body. Numerous cell experiment studies have successfully utilized microfluidic channels to achieve various new scientific discoveries. However, it has been often overlooked that undesired and unnoticed propagation of cellular molecules in such bio-microfluidic channel systems can have a negative impact on the experimental results. Thus, more careful designing is required to minimize such unwanted issues through deeper understanding and careful control of chemically and physically predominant factors at the microscopic scale. In this paper, we introduce a new approach to improve microfluidic channel design, specifically targeting the mitigation of the aforementioned challenges. To minimize the occurrence of undesired cell positioning upstream from the main test section where a concentration gradient field locates, an additional narrow port structure was devised between the microfluidic upstream channel and each inlet reservoir. This port also functioned as a passive lock that hold the flow at rest via fluid-air surface tension, which facilitated manual movement of the device even when cell attachment was not achieved completely. To demonstrate the practicability of the system, we conducted experiments and diffusion simulations on the effect of endocrine disruptors on germ cells. To this end, a bisphenol-A (BPA) concentration gradient was generated in the main channel of the system at BPA concentrations ranging from 120.8 μM to 79.3 μM, and the proliferation of GC-1 cells in the BPA gradient environment was quantitatively evaluated. The features and concepts of the introduced design is to minimize unexpected and ignored error sources, which will be one of the issues to be considered in the development of microfluidic systems to explore extremely delicate cellular phenomena.

A Miniaturized System for Rapid, Isothermal Detection of SARS-CoV-2 in Human and Environmental Samples.

We report a small-footprint cost-effective isothermal rapid DNA amplification system, with integrated microfluidics for automated sample analysis and detection of SARS-CoV-2 in human and environmental samples. Our system measures low-level fluorescent signals in real-time during amplification, while maintaining the desired assay temperature on a low power, portable system footprint. A unique soft microfluidic chip design was implemented to mitigate thermocapillary effects and facilitate optical alignment for automated image capture and signal analysis. The system-on-board prototype, coupled with the LAMP primers designed by BioCoS, was sensitive enough to detect large variations in viral loads of SARS-CoV-2 corresponding to a threshold cycle range of 16 to 39. Furthermore, tested samples consisted of a broad range of viral strains and lineages identified in Canada during 2021-2022. Clinical specimens were collected and tested at the Kingston Health Science Centre using a clinically validated PCR assay, and variants were determined using whole genome sequencing.

Towards a microfluidic H295R steroidogenesis assay-biocompatibility study and steroid detection on a thiol-ene-based chip.

The development of cell-based microfluidic assays offers exciting new opportunities in toxicity testing, allowing for integration of new functionalities, automation, and high throughput in comparison to traditional well-plate assays. As endocrine disruption caused by environmental chemicals and pharmaceuticals represents a growing global health burden, the purpose of the current study was to contribute towards the miniaturization of the H295R steroidogenesis assay, from the well-plate to the microfluidic format. Microfluidic chip fabrication with the established well-plate material polystyrene (PS) is expensive and complicated; PDMS and thiol-ene were therefore tested as potential chip materials for microfluidic H295R cell culture, and evaluated in terms of cell attachment, cell viability, and steroid synthesis in the absence and presence of collagen surface modification. Additionally, spike-recovery experiments were performed, to investigate potential steroid adsorption to chip materials. Cell aggregation with poor steroid recoveries was observed for PDMS, while cells formed monolayer cultures on the thiol-ene chip material, with cell viability and steroid synthesis comparable to cells grown on a PS surface. As thiol-ene overall displayed more favorable properties for H295R cell culture, a microfluidic chip design and corresponding cell seeding procedure were successfully developed, achieving repeatable and uniform cell distribution in microfluidic channels. Finally, H295R perfusion culture on thiol-ene chips was investigated at different flow rates (20, 10, and 2.5 µL/min), and 13 steroids were detected in eluting cell medium over 48h at the lowest flow rate. The presented work and results pave the way for a time-resolved microfluidic H295R steroidogenesis assay.

Few-body hydrodynamic interactions probed by optical trap pulling experiment.

We study the hydrodynamic coupling of neighboring micro-beads placed in a multiple optical trap setup allowing us to precisely control the degree of coupling and directly measure time-dependent trajectories of entrained beads. We performed measurements on configurations with increasing complexity starting with a pair of entrained beads moving in one dimension, then in two dimensions, and finally a triplet of beads moving in two dimensions. The average experimental trajectories of a probe bead compare well with the theoretical computation, illustrating the role of viscous coupling and setting timescales for probe bead relaxation. The findings also provide direct experimental corroborations of hydrodynamic coupling at large, micrometer spatial scales and long, millisecond timescales, of relevance to, e.g., microfluidic device design and hydrodynamic-assisted colloidal assembly, improving the capability of optical tweezers, and understanding the coupling between micrometer-scale objects within a living cell.

Expanding the toolbox for microfluidic-based in situ membrane characterization via microscopy

Polyamide thin film composite membranes are the commercial standard for aqueous nanofiltration and reverse osmosis. Establishing their synthesis-structure-performance relationships (SSPs), needed for rational membrane design, is hampered by the small scale and high reaction rate of interfacial polymerization (IP). Microfluidic devices, compatible with microscopic real-time visualization of IP and performance testing of the formed film, are interesting within this respect. In this study, a new microfluidic design and operational protocol for in situ characterization of IP is developed. Difficulties encountered with microfluidics and coping strategies are highlighted. The outcome of the optimization study proves that a parylene-coated PDMS-glass chip comprising a channel lay-out with 4 inlets, 2 outlets, a channel height of 20 μm, and a reaction channel length ≤50 μm is most compatible with IP and performance testing. Varying synthesis conditions show changing film morphology and water flux in line with trends for dip-coated membranes. Addition of NaHCO3 and ethyl acetate induce morphological features and increase water flux. Increasing TMC concentrations decrease water flux until an excess is generated. By combining the developed protocol and microfluidic device with an online measurement technique to probe film formation dynamics, such as fluorescence microscopy, SSPs can be derived in the future.