Fabrication Of Microfluidic Chips Research Articles

In situ counter-diffusion crystallization and long-term crystal preservation in microfluidic fixed targets for serial crystallography.

Compared with batch and vapor diffusion methods, counter diffusion can generate larger and higher-quality protein crystals yielding improved diffraction data and higher-resolution structures. Typically, counter-diffusion experiments are conducted in elongated chambers, such as glass capillaries, and the crystals are either directly measured in the capillary or extracted and mounted at the X-ray beamline. Despite the advantages of counter-diffusion protein crystallization, there are few fixed-target devices that utilize counter diffusion for crystallization. In this article, different designs of user-friendly counter-diffusion chambers are presented which can be used to grow large protein crystals in a 2D polymer microfluidic fixed-target chip. Methods for rapid chip fabrication using commercially available thin-film materials such as Mylar, propyl-ene and Kapton are also detailed. Rules of thumb are provided to tune the nucleation and crystal growth to meet users' needs while minimizing sample consumption. These designs provide a reliable approach to forming large crystals and maintaining their hydration for weeks and even months. This allows ample time to grow, select and preserve the best crystal batches before X-ray beam time. Importantly, the fixed-target microfluidic chip has a low background scatter and can be directly used at beamlines without any crystal handling, enabling crystal quality to be preserved. The approach is demonstrated with serial diffraction of photoactive yellow protein, yielding 1.32 Å resolution at room temperature. Fabrication of this standard microfluidic chip with commercially available thin films greatly simplifies fabrication and provides enhanced stability under vacuum. These advances will further broaden microfluidic fixed-target utilization by crystallographers.

The design and fabrication of thermoplastic microfluidic chips with integrated micropillars for particle separation

Microfluidic technology utilizing the deterministic lateral displacement (DLD) method holds significant promise for efficiently separating micro-particles and biological cells. Despite the notable high throughput advantages associated with DLD chips, their widespread application is impeded by the substantial manufacturing costs of tens of thousands of micropillars. This study aims to explore the feasibility of employing the injection molding method for the mass production of DLD microfluidic chips. A multistage DLD chip with varied critical diameters was designed to isolate white blood cells from the human whole blood. The separation effectiveness was verified with the polydimethylsiloxane chip fabricated by standard soft lithography. Subsequently, nickel mold inserts were electroformed to fabricate thermoplastic DLD chips via the injection molding. The replication quality of micropillars under different molding parameters was studied. The capability of injection-molded chips to effectively achieve particle separation was validated. Results showed that thermoplastic chips with good replication quality were obtained, providing a scale-up production strategy for fabricating polymer-based microfluidic chips for disposable separation applications.

Atmospheric pressure atomic layer deposition for in-channel surface modification of PDMS microfluidic chips

Polydimethylsiloxane (PDMS) is one of the materials of choice for the fabrication of microfluidic chips. However, its broad application is constrained by its incompatibility with common organic solvents and the absence of surface anchoring groups for surface functionalization. Current solutions involving bulk-, ex-situ surface-, and in-situ liquid phase modifications are limited and practically demanding. In this work, we present a simple, novel strategy to deposit a metal oxide nano-layer on the inside of bonded PDMS microfluidic channels using atmospheric pressure atomic layer deposition (AP-ALD). Using three important classes of microfluidic experiments, i.e., (i) the production of micron-sized particles, (ii) the cultivation of biological cells, and (iii) the photocatalytic degradation in continuous flow chemistry, we demonstrate that the metal oxide nano-layer offers a higher resistance against organic solvent swelling, higher hydrophilicity, and a higher degree of further functionalization of the wall. We demonstrate the versatility of the approach by not only depositing SiOx nano-layers, but also TiOx nano-layers, which in the case of the flow chemistry experiment were further functionalized with gold nanoparticles through the use of AP-ALD. This study demonstrates AP-ALD as a tool to broaden the applicability of PDMS devices.

Rapid fabrication of modular 3D paper-based microfluidic chips using projection-based 3D printing

Rapid fabrication of modular 3D paper-based microfluidic chips using projection-based 3D printing

Durable superhydrophobic LTA-zeolite coating on PDMS surface with excellent self-cleaning property

Superhydrophobicity is a fundamental characteristic that plays a vital role in various applications, such as self-cleaning coatings, water vapor barriers for flexible electronics, and microfluidic chip fabrication. In this study, we developed a superhydrophobic surface coating on polydimethylsiloxane (PDMS) using Linde Type A (LTA) zeolite crystals, employing a silane coupling strategy. As part of this study, the LTA-zeolite-coated PDMS surface is characterized using various techniques to elucidate its superhydrophobic behavior, and a functionalization mechanism is proposed. The resulting substrate exhibited high surface roughness, with a water contact angle exceeding 155°, a sliding angle below 1°, and a contact angle hysteresis under 10°. This superhydrophobic surface coating effectively repelled water, confirming its excellent self-cleaning properties. Furthermore, the coating withstood three cycles of the Kapton adhesive tape peeling test, 5 h of water-jet hitting, and 72 h of UV light exposure, demonstrating its robustness and strong adhesion to PDMS.

An Alternative Micro-Milling Fabrication Process for Rapid and Low-Cost Microfluidics.

Microfluidics is an important technology for the biomedical industry and is often utilised in our daily lives. Recent advances in micro-milling technology have allowed for rapid fabrication of smaller and more complex structures, at lower costs, making it a viable alternative to other fabrication methods. The microfluidic chip fabrication developed in this research is a step-by-step process with a self-contained wet milling chamber. Additionally, ethanol solvent bonding is used to allow microfluidic chips to be fully fabricated within approximately an hour. The effect of using this process is tested with quantitative contact profileometery data to determine the expected surface roughness in the microchannels. The effect of surface roughness on the controllability of microparticles is tested in functional microfluidic chips using image processing to calculate particle velocity. This process can produce high-quality channels when compared with similar studies in the literature and surface roughness affects the control of microparticles. Lastly, we discuss how the outcomes of this research can produce rapid and higher-quality microfluidic devices, leading to improvement in the research and development process within the fields of science that utilise microfluidic technology. Such as medicine, biology, chemistry, ecology, and aerospace.

Elucidating the mechanism of overcuring in microchannels fabricated via vat photopolymerization (VPP) for precise microfluidic chip printing

Vat Photopolymerization (VPP) technology is highly regarded for precision micromachining applications like microfluidic chip fabrication, owing to its fine resolution, adaptability, and rapid prototyping capabilities. However, the challenge of resin over-curing resulting from UV energy penetration poses a significant obstacle in microchannel printing. This study addresses this challenge by investigating the intricate relationship between resin properties and over-curing. Diverse microfluidic chips were printed utilizing VPP technology, and the overall cross-sectional over-curing characteristics of the channels were systematically examined. A novel combination of the classical UV light curing model and computational Fluid-Structure Interaction (FSI) model with the VPP printing process was proposed to examine the complex hydrodynamics and exfoliation dynamics within VPP-printed microchannels. The simulation results suggest that resins with higher viscosities and surface roughness are better suited for microchannel printing. Subsequent experiments using five different resins verified these findings and showed that the microchannels obtained with resins of lower viscosity and surface roughness resins have an irregular trapezoidal cross-section, which is uncontrollable and unpredictable and seriously affects the microchannel printing accuracy. Conversely, resins with higher viscosity and surface roughness have an ideal rectangular cross-section, which can be compensated by the model further refined printing to print microchannel. Finally, microfluidic chips were printed using "high precision 8 K resin" and "8 K-clay resin" with higher viscosity and surface roughness. The results proved that the microchannels printed by these resins had the desired rectangular cross-section, which was consistent with the "DLP light-curing" model. This study elucidates, for the first time, the intricate interplay between resin properties, printing structure, and microchannel over-curing, and this provides a solid groundwork for advancing high-precision microfluidic chip fabrication via DLP printing.

Fabrication of a Three-Dimensional Microfluidic System from Poly(methyl methacrylate) (PMMA) Using an Intermiscibility Vacuum Bonding Technique.

In this study, the fabrication of microfluidic chips through the bonding of poly (methyl methacrylate) (PMMA) boards featuring designed patterns to create a three-dimensional sandwich structure with embedded microchannels was explored. A key focus was optimization of the interface quality of bonded PMMA pairs by adjusting the solvent, such as such as acetone, alcohol, and their mixture. Annealing was conducted below 50 °C to leverage the advantages of low-temperature bonding. Because of the differences in the chemical reactivity of PMMA toward acetone, alcohol, and their combinations, the resulting defect densities at the bonding interfaces differed significantly under low-temperature annealing conditions. To achieve the optimal sealing integrity, bonding pressures of 30 N, 40 N, and 50 N were evaluated. The interface was analyzed through microstructural examination via optical microscopy and stress measurements were determined using digital photoelasticity, while the bonding strength was assessed through tensile testing.

Achieving High-Precision, Low-Cost Microfluidic Chip Fabrication with Flexible PCB Technology.

Soft lithography has long remained the state of the art to generate the necessary micropatterning for molded microfluidic (MF) chips. Previous attempts to use printed circuit boards (PCBs) as a cheap and accessible alternative to expensive lithographed molds for the production of PDMS MF chip prototypes have shown their limitations. A more in-depth exploration of using PCBs as a mold substrate and a novel methodology of using flexible PCBs to produce highly accurate MF chips is reported here for the first time. Cross sections highlight the improved accuracy of this method, and peel testing is performed to demonstrate suitable adhesion between the glass substrate and PDMS cast. Positive cell growth viability showcases this novel method as a high-accuracy, high-accessibility, low-cost prototyping method for microfluidic chips while still maintaining all favorable properties provided by the PDMS material.

Ultraviolet laser-etched Norland optical adhesive 81 micromodel for studying two-phase flow experiments

As the global energy demand grows, maximizing oil extraction from known reserves has become critical. The study of microfluidic flow and transport in porous media has become a key direction for future subsurface energy technologies. However, the high requirements of fabrication techniques and materials have constrained the progress of micro-scale experiments. In this study, we have innovatively proposed a microfluidic chip fabrication method based on ultraviolet laser, and a set of visualized microdrive platforms is developed to allow direct observation of two-phase flow processes at the pore scale. In this study, two pore structures—one with low porosity and high connectivity and the other with high porosity but low connectivity—were constructed to investigate the effect of pore structure on recovery. Two micromodels with different pore structures were fabricated, and water and surfactant drive experiments were conducted at different drive rates, respectively. The results show that increasing the replacement rate and introducing surfactant can somewhat improve the final recovery. Using surfactant is more effective in enhancing the recovery rate than increasing the replacement rate. The complexity of pore structure is one of the main factors affecting the formation of residual oil. The microfluidic experimental setup proposed in this study reduces the time and cost of conventional practical methods. It permits visualization of the oil drive process, demonstrating that the Norland Optical Adhesive 81 (NOA81) micromodel is a valuable tool in two-phase flow studies and its applications.

TPP-Based Microfluidic Chip Design and Fabrication Method for Optimized Nerve Cells Directed Growth

Microfluidic chips offer high customizability and excellent biocompatibility, holding important promise for the precise control of biological growth at the microscale. However, the microfluidic chips employed in the studies of regulating cell growth are typically fabricated through 2D photolithography. This approach partially restricts the diversity of cell growth platform designs and manufacturing efficiency. This paper presents a method for designing and manufacturing neural cell culture microfluidic chips (NCMC) using two-photon polymerization (TPP), where the discrete and directional cell growth is optimized through studying the associated geometric parameters of on-chip microchannels. This study involves simulations and discussions regarding the effects of different hatching distances on the mold surface topography and printing time in the Describe print preview module, which determines the appropriate printing accuracy corresponding to the desired mold structure. With the assistance of the 3D maskless lithography system, micron-level rapid printing of target molds with different dimensions were achieved. For NCMC with different geometric parameters, COMSOL software was used to simulate the local flow velocity and shear stress characteristics within the microchannels. SH-SY5Y cells were selected for directional differentiation experiments on NCMC with different geometric parameters. The results demonstrate that the TPP-based manufacturing method efficiently constructs neural microfluidic chips with high precision, optimizing the discrete and directional cell growth. We anticipate that our method for designing and manufacturing NCMC will hold great promise in construction and application of microscale 3D drug models.

Microfluidic Chip Fabrication for Tumor Cell 3D Culture Based on Microwell Arrays.

Compared with traditional 2D cell culture, 3D cell culture more closely resembles the original state of cells in vivo and enables the establishment of in vivo-like microenvironments and cell-cell interactions, thereby providing valuable cellular materials for numerous studies. The direct establishment of in vitro patient tumor models can enhance drug testing, cancer research, and individualized precision therapy. In this study, we propose a microfluidic chip based on microwell arrays for 3D tumor cell culture. This chip combines nanoscale channels and microwell arrays to precisely control cell distribution and nutrient diffusion, thus closely mimicking the tumor microenvironment. The incorporation of microwell arrays allows for simple and rapid high-throughput preparation of tumor spheroids, while promoting the formation of cell-cell and cell-matrix interactions, ultimately enhancing cell viability and function. Preliminary experiments using tumor cell lines validate the ability of the chip to support 3D tumor growth with enhanced physiological relevance. The microfluidic chip serves as a reliable and scalable platform for studying tumor biology and evaluating therapeutic efficacy and is anticipated to expedite cancer research and drug discovery.

Micro‐Patterning Wettability in Very Large Scale Microfluidic Integrated Chips for Double Emulsion Generation

AbstractThe fabrication of microfluidic chips is becoming a mature field and channels can be reliably defined with micrometer‐scale precision in a variety of substrates. In addition to channel geometry, micrometer‐scale patterning of wettability in microfluidic channels is essential for many applications including multi‐phase flow stabilization and multiple emulsion generation. Unfortunately, current methods to pattern wettability in microfluidics suffer from low spatial resolution, inability for patterns to be arbitrarily defined, cumbersome procedures, and incompatibility with parallelized architectures for scaled‐up production of microfluidic generated materials. To address these issues, a method is developed to lithographically define micrometer‐scale resolution patterns of wettability on all channel surfaces (ceiling, floor, and walls) in silicon and glass microfluidic devices with complex 3D geometry. A process is reported to pattern silanes on microfluidic chips that uses photolithography and an optimized process that keeps silanized surfaces stable through the microfabrication process, including anodic bonding. The versatility of this approach is highlighted by patterning wettability of a silicon/glass device to generate both highly uniform water–in‐oil–in‐water and oil–in‐water–in‐oil double emulsions. The applicability of this process is demonstrated to the parallel generation of materials in a microfluidic chip with complex geometry, by fabricating and successfully validating parallelized double emulsion generators.

A mould-free soft-lithography approach for rapid, low-cost and bulk fabrication of microfluidic chips using photopolymer sheets

A mould-free soft-lithography approach for rapid, low-cost and bulk fabrication of microfluidic chips using photopolymer sheets

Engineering Cardiac Tissue for Advanced Heart-On-A-Chip Platforms.

Cardiovascular disease is a major cause of mortality worldwide, and current preclinical models including traditional animal models and 2D cell culture models have limitations in replicating human native heart physiology and response to drugs. Heart-on-a-chip (HoC) technology offers a promising solution by combining the advantages of cardiac tissue engineering and microfluidics to create in vitro 3D cardiac models, which can mimic key aspects of human microphysiological systems and provide controllable microenvironments. Herein, recent advances in HoC technologies are introduced, including engineered cardiac microtissue construction in vitro, microfluidic chip fabrication, microenvironmental stimulation, and real-time feedback systems. The development of cardiac tissue engineering methods is focused for 3D microtissue preparation, advanced strategies for HoC fabrication, and current applications of these platforms. Major challenges in HoC fabrication are discussed and the perspective on the potential for these platforms is provided to advance research and clinical applications.

Fabrication and Validation of a 3D Portable PEGDA Microfluidic Chip for Visual Colorimetric Detection of Captured Breast Cancer Cells.

To guide therapeutic strategies and to monitor the state changes in the disease, a low-cost, portable, and easily fabricated microfluidic-chip-integrated three-dimensional (3D) microchamber was designed for capturing and analyzing breast cancer cells. Optimally, a colorimetric sensor array was integrated into a microfluidic chip to discriminate the metabolites of the cells. The ultraviolet polymerization characteristic of poly(ethylene glycol) diacrylate (PEGDA) hydrogel was utilized to rapidly fabricate a three-layer hydrogel microfluidic chip with the designed structure under noninvasive 365 nm laser irradiation. 2-Hydroxyethyl methacrylate (HEMA) was added to the prepolymer in order to increase the adhesive capacity of the microchip's surface for capturing cells. 1-Vinyl-2-pyrrolidone (NVP) was designed to improve the toughness and reduce the swelling capacity of the hydrogel composite. A non-toxic 3D hydrogel microarray chip (60 mm × 20 mm × 3 mm) with low immunogenicity and high hydrophilicity was created to simulate the real physiological microenvironment of breast tissue. The crisscross channels were designed to ensure homogeneous seeding density. This hydrogel material displayed excellent biocompatibility and tunable physical properties compared with traditional microfluidic chip materials and can be directly processed to obtain the most desirable microstructure. The feasibility of using a PEGDA hydrogel microfluidic chip for the real-time online detection of breast cancer cells' metabolism was confirmed using a specifically designed colorimetric sensor array with 16 kinds of porphyrin, porphyrin derivatives, and indicator dyes. The results of the principal component analysis (PCA), the hierarchical cluster analysis (HCA), and the linear discriminant analysis (LDA) suggest that the metabolic liquids of different breast cells can be easily distinguished with the developed PEGDA hydrogel microfluidic chip. The PEGDA hydrogel microfluidic chip has potential practicable applicability in distinguishing normal and cancerous breast cells.

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.

Fabrication of Spiral Low-Cost Microchannel with Trapezoidal Cross Section for Cell Separation Using a Grayscale Approach.

Trapezoidal cross-sectional spiral microfluidic channels showed high resolution and throughput in cell separation in bio-applications. The main challenges are the complexity and high cost of the fabrication process of trapezoidal cross-sectional channels on the micro-scale. In this work, we present the application of grayscale in microfluidic channel design to overcome the complexity of the fabrication process. We also use direct engraving with a CO2 laser beam on polymethyl methacrylate (PMMA) material to drastically reduce the microfluidic chip's cost (to <30 cents) and fabrication time (to 20 min). The capability of the present fabrication methodology for cell sorting applications is demonstrated through experimental tests for the separation of white blood cells (WBCs) from whole blood at different dilution factors. The experimental results indicated that an 800 µL/min flow rate provided the optimal separation efficiency using the fabricated chip. A 90.14% separation efficiency at 1% hematocrit diluted blood sample was reported.

Design and Fabrication of a High Performance Microfluidic Chip for Blood Plasma Separation: Modelling and Prediction of System Behaviour via CFD Method.

This paper presents a single-step microfluidic system designed for passive separation of human fresh blood plasma using direct capillary forces. Our microfluidic system is composed of a cylindrical well between upper and lower channel pairs produced by soft photolithography. The microchip was fabricated based on hydrophobicity differences upon suitable cylindrical surfaces using gravitational and capillary forces and lateral migration of plasma and red blood cells. The plasma radiation was applied to attach the polymeric segment (polydimethylsiloxane (PDMS)) to the glass. Meanwhile, Tween 80 was used as a surfactant to increase the hydrophobicity of the lateral channel surfaces. This led to the higher movement of whole blood, including plasma. Fick's law of diffusion was validated for this diffusion transfer, the Navier-Stokes equation was used for the momentum balance, and the Laplace equation was utilized for the dynamics of the mesh. A model with high accuracy using the COMSOL Multiphysics software was created to predict the capillary forces and chip model validation. RBCs (red blood cells) were measured by the H3 cell counter instrument, by which 99% plasma purity was achieved. Practically, 58.3% of the plasma was separated from the blood within 12 min. Correlation between plasma separation results obtained from software and experimental data showed a coefficient of determination equal to 0.9732. This simple, rapid, stable, and reliable microchip can be considered as a promising candidate for providing plasma in point-of-care diagnostics.

Research on Integrated 3D Printing of Microfluidic Chips.

Microfluidic chips have the advantages of miniaturization, integration, and portability, and are widely used in the early diagnosis of major diseases, personalized medical treatment, environmental detection, health quarantine, and other fields. The existing microfluidic chip manufacturing process is difficult to operate because of complex three-dimensional channels, complicated manufacturing steps, limited printing materials, the difficulty of operating the bonding process, and the need to purchase expensive new equipment. In this paper, an integrated molding method for microfluidic chips that integrates 3D printing and polymer dissolution technology is proposed. First, the channel mold of poly(vinyl alcohol) (PVA) or high impact polystyrene (HIPS) is dissolved to complete the manufacturing of the microfluidic chip channel. The integrated 3D-forming method of microfluidic chips proposed in this paper can manufacture microchannels inside the microfluidic chip, avoid the bonding process, and eliminate the need for rapid alignment of microchannels, material modification, and other operations, thus improving the stability of the process. Finally, by comparing the microchannels made by PVA and HIPS, it is concluded that the quality of the microchannels made by HIPS is obviously better than that made by PVA. This paper provides a new idea for the fabrication of microfluidic chips and the application of HIPS.