Fabrication Of Microfluidic Devices Research Articles

Development of a direct PMMA-PCB bonding method for low cost and rapid prototyping of microfluidic-based gas analysers.

Rapid prototyping of microfluidic devices requires low cost materials and simple fabrication methods. PMMA and PCB have been used separately for the fabrication of microfluidic devices in a wide range of applications. Although the combined use of PMMA and PCB can have considerable merits, few works have been reported on the direct bonding of these materials. In this work we have investigated the fabrication of microfluidic devices using PMMA and PCB for the analysis of gaseous samples. In order to yield a reliable direct bonding method, four parameters including temperature, pressure, solvent and patterned interface material were experimentally investigated. Results of testing various prototypes showed that a patterned interface of concentric rectangular copper rings exposed to solvent at room temperature and under moderate pressure provided better adhesion strength, sealing and durability. After successful development of the PMMA-PCB direct bonding process, sample prototypes were designed and fabricated to practically assess the combined advantages of two materials. Presented concepts include implementation of heater on a PCB, array of gas sensors coupled with microchannels, serpentine microchannel and fast evaporation of liquid sample using an SMD resistor. It has been shown that advantages of utilizing PMMA such as fabricating the channel easily and with low cost, can be combined with benefits of a PCB including simple sensor installation and the use of copper tracks and electronic components for gas flow modulation. Moreover, it is possible to implement channel, circuit and other electronic components such as microprocessors on a single device.

Single-step batch fabrication of microfluidic paper-based analytical devices with a 3D printer and their applications in nanoenzyme-enhanced visual detection of dopamine.

Wax printing is the most widely used method for fabricating microfluidic paper-based analytical devices (μPADs), but it still suffers from disadvantages like discontinuation of wax printers and need for additional equipment for heating treatment. To address these issues, this work initially describes a new class of wax printing approach for high-precision, batch fabrication of μPADs using a household 3D printer. It only involves a one patterning step of printing polyethylene wax into rice paper body. Under optimized parameters, a fabrication resolution, namely the minimum hydrophilic channel width, down to ~189 ± 30 μm could be achieved. In addition, the analytical applicability of such polyethylene wax-patterned μPADs was demonstrated well with enhanced colorimetric detection of dopamine as a model analyte by combining metal-organic framework (MOF) based nanoenzymes (ZIF-67) with a smartphone (for portable quantitative readout). The developed nanosensor could linearly detect dopamine over a concentration range from 10 to 1000 μM, with a detection limit of ca. 2.75 μM (3σ). The recovery results for analyzing several real samples (i.e., pig feed, chicken feed, pork and human serum) were between 91.82 and 102.79%, further validating its good detection accuracy for potential practical applications in food safety and medical diagnosis.

Validation of Fluid Flow Speed Behavior in Capillary Microchannels Using Additive Manufacturing (SLA Technology)

This research explores fluid flow speed behavior in capillary channels using additive manufacturing, focusing on stereolithography (SLA). It aims to validate microchannels fabricated through SLA for desired fluid flow characteristics, particularly capillary-driven flow. The methodology involves designing, fabricating, and characterizing microchannels via SLA, with improvements such as an air-cleaning step facilitating the production of microchannels ranging from 300 to 1000 µm. Experimental validation assesses fluid flow speed behavior across channels of varying dimensions, evaluating the impact of channel geometry, surface roughness, and manufacturing parameters. The findings affirm the feasibility and efficacy of SLA in producing microchannels with consistent and predictable fluid flow behavior between 300 to 800 µm. This study contributes insights into microfluidic device fabrication techniques and enhances the understanding of fluid dynamics in capillary-driven systems. Overall, it underscores the potential of additive manufacturing, specifically SLA, in offering cost-effective and scalable solutions for microfluidic applications. The validated fluid flow speed behavior in capillary channels suggests new avenues for developing innovative microfluidic devices with improved performance and functionality, marking a significant advancement in the field.

Saturation Equation: An Analytical Expression for Partial Saturation during Wicking Flow in Paper Microfluidic Channels.

The design and fabrication of paper-based microfluidic devices is critically dependent on modeling fluid flow through porous paper membranes. A commonly observed phenomenon is partial saturation, i.e., regions of the paper membrane not being filled completely due to pores of different sizes. The most comprehensive model to date of partial saturation during wicking flow in paper is the Richards equation. However, the solution to the Richards equation requires numerical solvers like COMSOL, which makes it largely inaccessible to the paper microfluidics and lateral flow assay community. There is therefore a need for a simple and appropriate model of partial saturation in paper membranes, easily usable by the wider research community. In the current work, we present an approach to model paper membranes as a bundle of parallel capillaries whose radii follow a two-parameter log-normal distribution. Application of the Washburn equation to the bundle provides a distribution of fluid fronts, which can be used to calculate saturation. Using this approach, we developed the "saturation equation"─an explicit analytical expression to calculate saturation as a function of space and time in 1D wicking flow. Experimentally obtained data for spatiotemporal saturation for four different paper materials were fit to this analytical model to obtain parameters for each material. Results obtained from this analytical model match well with both experimental data and numerical results obtained from the Richards equation. The availability of an explicit analytical expression for partial saturation will enable incorporation of the critical phenomenon of partial saturation in the design of paper microfluidic devices.

Double-Sided Tape in Microfluidics: A Cost-Effective Method in Device Fabrication.

The demand for easy-to-use, affordable, accessible, and reliable technology is increasing in biological, chemical, and medical research. Microfluidic devices have the potential to meet these standards by offering cost-effective, highly sensitive, and highly specific diagnostic tests with rapid performance and minimal sample volumes. Traditional microfluidic device fabrication methods, such as photolithography and soft lithography, are time-consuming and require specialized equipment and expertise, making them costly and less accessible to researchers and clinicians and limiting the applicability and potential of microfluidic devices. To address this, researchers have turned to using new low-cost materials, such as double-sided tape for microfluidic device fabrication, which offers simple and low-cost processes. The innovation of low-cost and easy-to-make microfluidic devices improves the potential for more devices to be transitioned from laboratories to commercialized products found in stores, offices, and homes. This review serves as a comprehensive summary of the growing interest in and use of double-sided tape-based microfluidic devices in the last 20 years. It discusses the advantages of using double-sided tape, the fabrication techniques used to create and bond microfluidic devices, and the limitations of this approach in certain applications.

Two‐Photon Direct Laser Writing of pNIPAM Actuators in Microchannels for Dynamic Microfluidics

Microfluidic tools enable to investigate and manipulate various chemical and biological processes at small scales. As a result, it finds widespread applications in lab‐on‐chip devices, drug delivery systems, or miniaturized cell cultures. However, microfluidic devices are still limited in their flexibility and are often designed to fulfill a single functionality. Moreover, technologies to introduce dynamic functionalities with high precision and at high resolution after the development of a continuous phase microfluidic chip remain scarce. Herein, two‐photon polymerization direct laser writing is introduced as a suitable approach to equip continuous phase microfluidic chips with structurally defined thermoresponsive poly(N‐isopropyl‐acrylamide) (pNIPAM) microactuators. Harnessing the lower critical phase transition temperature of pNIPAM, and upon controlling specific design parameters, the efficient catch and release of polystyrene beads of different sizes using a pNIPAM micropillar brush array is demonstrated. Moreover, a biocompatible pNIPAM microgripper array is designed to subsequently capture and release differently sized (single) cell populations. Overall, the method offers great flexibility and a high degree of freedom toward the fabrication of dynamic microfluidic devices with great adaptability to experimental conditions in real time.

Effect of Mold Materials Used During Hot Embossing on Feature Fidelity for Microfabrication in Cyclic Olefin Polymer (COP) Substrate

During the transition from research to market, the fabrication of microfluidic devices in thermoplastic substrates is inevitable. For short production runs of several hundred products, hot embossing is the typical method before moving on to a typically more expensive injection molding process for higher production volumes. In this work, we investigated the effect of mold material used during hot embossing on feature fidelity for microfabrication in cyclic olefin polymer (COP) substrate. Specifically, we designed a simple flow-focusing microfluidic device and fabricated three different molds using silicon wafer by deep reactive ion etching (DRIE), aluminum filled high temperature epoxy by soft lithography and aluminum by CNC milling. We performed hot embossing experiments with 2mm thick COP substrates and these three different molds using automatic bench top Carver hot press. Finally, we characterized the hot embossed substrates by optical and scanning electron microscopy. Fabrication results demonstrate that the mold material plays a big role in feature fidelity. Among the mold materials used, silicon substrate performed the worst based on defects after demolding. Epoxy and aluminum molds were similar in terms of microfabricated feature defects in the substrate which could be mostly attributed to their coefficient of thermal expansion (CTE). A mold material with a CTE closer to the thermoplastic will result in much better feature fidelity.

Evolution of 3d printing technology in fabrication of microfluidic devices and biological applications: a comprehensive review

Lab-on-a-chip or LOC is a term that is used to describe microfluidic devices that integrate multiple analyte detection, which are normally carried out in a laboratory, into one micro-chip unit and may have applications in diverse fields such as electronics, medicine and biomedical domains. Even though microfluidics has advanced greatly during the past decade due to increased needs for portability, reduced sample requirement and multiple analyte detection capabilities biological research has not adopted the technology at the required pace. This may be owing to the time-consuming and expensive process involved in the microfabrication of biochips, the requirement of specialised setup facilities and the extremely high cost associated with microfluidics as compared to conventional technologies. In recent years, three-dimensional (3D) printing has piqued curiosity in the scientific community. It has the potential to create complex, high-resolution structures and that too in a short timeframe depending upon device complexity. This could inspire progressive research in microfluidics, particularly finding applications in biomedical engineering and point-of-care diagnostics. This article gives an overview of how 3D printing aids in the manufacture of microfluidic devices for biological applications, as well as the existing 3D printing methods which are utilised for fabrication and the future perspective in the development of microfluidic devices.

ADVANCES IN ULTRAPRECISION DIAMOND TURNING: TECHNIQUES, APPLICATIONS, AND FUTURE TRENDS

Advances in ultraprecision diamond turning have revolutionized manufacturing processes across various industries, offering unparalleled precision and surface quality in the fabrication of optical components, microfluidic devices, and advanced mechanical parts. This review delves into the techniques, applications, and future trends in ultraprecision diamond turning, highlighting recent advancements and potential trajectories. Techniques in ultraprecision diamond turning have evolved significantly, driven by innovations in machine design, tooling materials, and control systems. Diamond turning machines equipped with ultra-stiff structures, high-speed spindles, and advanced feedback mechanisms enable sub-nanometer level accuracy and surface finishes down to Angstrom levels. Additionally, advancements in single-point diamond turning (SPDT), fast tool servo (FTS), and deterministic microgrinding (DMG) techniques further enhance the versatility and precision of the process. Applications of ultraprecision diamond turning span a wide range of industries, including aerospace, automotive, biomedical, and telecommunications. In optics manufacturing, diamond turning facilitates the production of aspheric lenses, freeform optics, and diffractive optical elements with unprecedented accuracy, contributing to the development of high-performance imaging systems and laser applications. Moreover, in the biomedical field, diamond-turned microfluidic devices enable precise control over fluid flow and particle manipulation, empowering advancements in drug delivery systems and lab-on-a-chip technologies. Future trends in ultraprecision diamond turning are poised to address challenges related to scalability, multi-material processing, and in-situ metrology. Integration of adaptive control algorithms and machine learning techniques promises enhanced process stability and predictive maintenance, optimizing productivity and reducing downtime. Furthermore, the development of hybrid manufacturing approaches, combining diamond turning with additive manufacturing or laser processing, offers novel avenues for fabricating complex, multi-functional components with improved efficiency and cost-effectiveness. The ongoing advancements in ultraprecision diamond turning techniques, coupled with diverse applications across industries, underscore its pivotal role in advancing manufacturing capabilities. Anticipated future trends hold promise for further expanding the scope and impact of this technology, driving innovation and pushing the boundaries of precision engineering.
 Keywords: Ultraprecision, Diamond, Turning, Technique, Review.

Beyond 2D: Novel biomaterial approaches for modeling the placenta

This review considers fully three-dimensional biomaterial environments of varying complexity as these pertain to research on the placenta. The developments in placental cell sources are first considered, along with the corresponding maternal cells with which the trophoblast interact. We consider biomaterial sources, including hybrid and composite biomaterials. Properties and characterization of biomaterials are discussed in the context of material design for specific placental applications. The development of increasingly complicated three-dimensional structures includes examples of advanced fabrication methods such as microfluidic device fabrication and 3D bioprinting, as utilized in a placenta context. The review finishes with a discussion of the potential for in vitro, three-dimensional placenta research to address health disparities and sexual dimorphism, especially in light of the exciting recent changes in the regulatory environment for in vitro devices.

Characterization of Shrink Film Properties for Rapid Microfluidics Lab-on-Chip Fabrication.

Shrink film is a thin sheet of polystyrene plastic that shrinks to 25-40% of its original size when heated. This study investigated the shrinkage factor of the film at different temperatures and baking times to determine the optimal fabrication recipe for shrink film microfluidic device production. Additionally, this study characterized the properties of shrink film, including minimum possible feature size and cross-section geometries, using manual engraving and the CAMEO 4 automated cutting machine. The optimal shrinkage factor ranged from 1.7 to 2.9 at 150 °C and a baking time of 4 min, producing the ideal size for microfluidic device fabrication. The X- and Y-axes shrank ~2.5 times, while Z-axis thickened by a factor of ~5.8 times. This study achieved a minimum feature size of 200 microns, limited by the collapsing of channel sidewalls when shrunk, leading to blockages in the microchannel. These findings demonstrate the feasibility and versatility of using shrink film as a cost-effective and efficient material for the rapid fabrication of microfluidic devices. The potential applications of this material in various fields such as the medical and biomedical industries, bacteria and algae culture and enumeration are noteworthy.

Enhancing fabrication of hybrid microfluidic devices through silane‐based bonding: A focus on polydimethylsiloxane‐cyclic olefin copolymer and PDMS‐lithium niobate

AbstractEffective manipulation and control of fluids in microfluidic channels requires robust bonding between the different components. Polydimethylsiloxane (PDMS) is widely employed in microchannel fabrication due to its affordability, biocompatibility, and straightforward fabrication process. However, PDMS's low surface energy poses challenges in bonding with many organic and inorganic substrates, hindering the development of hybrid microfluidic devices. In this study, a simple and versatile three step process is presented for bonding PDMS microchannels with organic (cyclic olefin copolymer (COC)) and inorganic substrates (lithium niobate (LiNbO3)) using plasma activation and a silane coupling agent. Initially, the PDMS surface undergoes oxygen/argon plasma activation, followed by functionalization with (3‐aminopropyl) triethoxysilane (APTES). Subsequently, the COC or LiNbO3 is plasma activated and brought into contact with PDMS under a load at a specific temperature. Characterization by Fourier transform infrared, scanning electron microscopy, atomic force microscopy, and contact angle measurements confirmed the successful treatment of the substrates. In addition, bonding strength of the fabricated hybrid devices was assessed through leakage and tensile tests. Under optimized conditions (100°C and 4% v/v APTES), PDMS‐COC hybrid microchannels achieved a flow rate of 600 mL/h without leakage and a tensile strength of 562 kPa. Conversely, the PDMS‐ LiNbO3 assembly demonstrated a flow rate of 216 mL/h before leakage, with a tensile strength of 334 kPa. This bonding method exhibits significant potential and versatility for various materials in microfluidic applications, ranging from biomedical research to enhanced oil recovery.

Optimizing laser-based micro-cutting for PMMA microfluidic device fabrication: thermal analysis and parameter optimization

Abstract Laser processes have gained popularity in microfluidic device fabrication. This study aims to determine the optimal parameters for laser-based micro-cutting to achieve the desired width, depth, profile, and material removal, considering the thermal properties of PMMA. A CNC CO2 laser was used, and 29 trials tested various speed and power combinations. Two theoretical models based on trial results focused on depth and width. COMSOL Multiphysics FEA software estimated surface temperature. Theoretical depth estimation matched experimental data more accurately when the P/S ratio was below 0.15 and the scanning speed was set at 500 mm/s or 750 mm/s. At 500 mm/s, width estimation was most accurate, up to 30 W. At 750 mm/s, experimental width exceeded predictions. Material removal increased proportionally with increasing P/S ratio, but beyond a threshold of 0.15, material removal remained nearly constant despite rising heat input. Laser-cut track shape varied, resembling a ‘U’ at lower and a ‘V’ at higher ratios. The groove shape transitioned from ‘U’ to ‘V’ when the temperature surpassed 1200 K. A V-shaped groove required a temperature exceeding 1500 K. Optimization confirmed a microchannel depth of 0.197 mm, width of 0.256 mm, and ‘U–V’ channel shape achievable at 30 W and 200 mm/s scanning speed, with a surface temperature of 1325 K.

High-resolution low-cost LCD 3D printing for microfluidics and organ-on-a-chip devices.

The fabrication of microfluidic devices has progressed from cleanroom manufacturing to replica molding in polymers, and more recently to direct manufacturing by subtractive (e.g., laser machining) and additive (e.g., 3D printing) techniques, notably digital light processing (DLP) photopolymerization. However, many methods require technical expertise and DLP 3D printers remain expensive at a cost ∼15-30 K USD with ∼8 M pixels that are 25-40 μm in size. Here, we introduce (i) the use of low-cost (∼150-600 USD) liquid crystal display (LCD) photopolymerization 3D printing with ∼8-58 M pixels that are 18-35 μm in size for direct microfluidic device fabrication, and (ii) a poly(ethylene glycol) diacrylate-based ink developed for LCD 3D printing (PLInk). We optimized PLInk for high resolution, fast 3D printing and biocompatibility while considering the illumination inhomogeneity and low power density of LCD 3D printers. We made lateral features as small as 75 μm, 22 μm-thick embedded membranes, and circular channels with a 110 μm radius. We 3D printed microfluidic devices previously manufactured by other methods, including an embedded 3D micromixer, a membrane microvalve, and an autonomous capillaric circuit (CC) deployed for interferon-γ detection with excellent performance (limit of detection: 12 pg mL-1, CV: 6.8%). We made PLInk-based organ-on-a-chip devices in 384-well plate format and produced 3420 individual devices within an 8 h print run. We used the devices to co-culture two spheroids separated by a vascular barrier over 5 days and observed endothelial sprouting, cellular reorganization, and migration. LCD 3D printing together with tailored inks pave the way for democratizing access to high-resolution manufacturing of ready-to-use microfluidic and organ-on-a-chip devices by anyone, anywhere.

Mechanical Compression of Drosophila Embryos Using Rapid Fabrication Microfluidic Devices.

Microfluidic devices support developmental and mechanobiology studies by enabling the precise control of electrical, chemical, and mechanical stimuli at the microscale. Here, we describe the fabrication of customizable microfluidic devices and demonstrate their efficacy in applying mechanical loads to micro-organs and whole organisms, such as Drosophila embryos. The fabrication technique consists in the use of xurography to define channels and chambers using thin layers of thermoplastics and glass. The superposition of layers followed by thermal lamination produces robust and reproducible devices that are easily adapted for a variety of experiments. The integration of deformable layers and glass in these devices facilitates the imaging of cellular and molecular dynamics in biological specimens under mechanical loads. The method is highly adaptable for studies in mechanobiology.

Aerosol jet printing of surface acoustic wave microfluidic devices

The addition of surface acoustic wave (SAW) technologies to microfluidics has greatly advanced lab-on-a-chip applications due to their unique and powerful attributes, including high-precision manipulation, versatility, integrability, biocompatibility, contactless nature, and rapid actuation. However, the development of SAW microfluidic devices is limited by complex and time-consuming micro/nanofabrication techniques and access to cleanroom facilities for multistep photolithography and vacuum-based processing. To simplify the fabrication of SAW microfluidic devices with customizable dimensions and functions, we utilized the additive manufacturing technique of aerosol jet printing. We successfully fabricated customized SAW microfluidic devices of varying materials, including silver nanowires, graphene, and poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS). To characterize and compare the acoustic actuation performance of these aerosol jet printed SAW microfluidic devices with their cleanroom-fabricated counterparts, the wave displacements and resonant frequencies of the different fabricated devices were directly measured through scanning laser Doppler vibrometry. Finally, to exhibit the capability of the aerosol jet printed devices for lab-on-a-chip applications, we successfully conducted acoustic streaming and particle concentration experiments. Overall, we demonstrated a novel solution-based, direct-write, single-step, cleanroom-free additive manufacturing technique to rapidly develop SAW microfluidic devices that shows viability for applications in the fields of biology, chemistry, engineering, and medicine.

Microfluidics: a concise review of the history, principles, design, applications, and future outlook.

Microfluidic technologies have garnered significant attention due to their ability to rapidly process samples and precisely manipulate fluids in assays, making them an attractive alternative to conventional experimental methods. With the potential for revolutionary capabilities in the future, this concise review provides readers with insights into the fascinating world of microfluidics. It begins by introducing the subject's historical background, allowing readers to familiarize themselves with the basics. The review then delves into the fundamental principles, discussing the underlying phenomena at play. Additionally, it highlights the different aspects of microfluidic device design, classification, and fabrication. Furthermore, the paper explores various applications, the global market, recent advancements, and challenges in the field. Finally, the review presents a positive outlook on trends and draws lessons to support the future flourishing of microfluidic technologies.

Integrated microfluidic platforms for heavy metal sensing: a comprehensive review.

Heavy metals are found naturally; however, anthropogenic activities such as mining, inappropriate disposal of industrial waste, and the use of pesticides and fertilizers containing heavy metals can cause their unwanted release into the environment. Conventionally, detection of heavy metals is performed using atomic absorption spectrometry, electrochemical methods and inductively coupled plasma-mass spectrometry; however, they involve expensive and sophisticated instruments and multistep sample preparation that require expertise for accurate results. In contrast, microfluidic devices involve rapid, cost-efficient, simple, and reliable approaches for in-laboratory and real-time monitoring of heavy metals. The use of inexpensive and environment friendly materials for fabrication of microfluidic devices has increased the manufacturing efficiency of the devices. Different types of techniques used in heavy metal detection include colorimetry, absorbance-based, and electrochemical detection. This review provides insight into the detection of toxic heavy metals such as mercury (Hg), cadmium (Cd), lead (Pb), and arsenic (As). Importance is given to colorimetry, optical, and electrochemical techniques applied for the detection of heavy metals using microfluidics and their modifications to improve the limit of detection (LOD).

Surface-Wetting Characteristics of DLP-Based 3D Printing Outcomes under Various Printing Conditions for Microfluidic Device Fabrication.

In recent times, the utilization of three-dimensional (3D) printing technology, particularly a variant using digital light processing (DLP), has gained increasing fascination in the realm of microfluidic research because it has proven advantageous and expedient for constructing microscale 3D structures. The surface wetting characteristics (e.g., contact angle and contact angle hysteresis) of 3D-printed microstructures are crucial factors influencing the operational effectiveness of 3D-printed microfluidic devices. Therefore, this study systematically examines the surface wetting characteristics of DLP-based 3D printing objects, focusing on various printing conditions such as lamination (or layer) thickness and direction. We preferentially examine the impact of lamination thickness on the surface roughness of 3D-printed structures through a quantitative assessment using a confocal laser scanning microscope. The influence of lamination thicknesses and lamination direction on the contact angle and contact angle hysteresis of both aqueous and oil droplets on the surfaces of 3D-printed outputs is then quantified. Finally, the performance of a DLP 3D-printed microfluidic device under various printing conditions is assessed. Current research indicates a connection between printing parameters, surface roughness, wetting properties, and capillary movement in 3D-printed microchannels. This correlation will greatly aid in the progress of microfluidic devices produced using DLP-based 3D printing technology.

Print-Pause-Print Fabrication of Tailored Electrochemical Microfluidic Devices.

Three-dimensional (3D) printing technology has emerged as a powerful technology for the fabrication of low-cost microfluidics. Nevertheless, the fabrication of microfluidic devices integrating high-performance electrochemical sensors in practical applications is still an open challenge. Although automatic fabrication of the microfluidic device and the electrodes can be successfully carried out using a one-step multimaterial fused filament fabrication (FFF) approach, the as-printed electrochemical performance of these electrodes is not good enough for chemical (bio)sensing and their surface modification is challenging because after closing the channel there is no physical access to the electrode. Thus, here a pause-print-pause (PPP) microfabrication approach was implemented. The fabrication was paused before printing the microfluidics, and the filament-based electrodes were directly modified on the printing bed via stencil printing, drop casting, and electrodeposition. To exemplify this versatile workflow, the design of a microfluidic glucose sensor was proposed. To this end, first, the working and counter electrodes were stencil printed with graphite ink while the reference electrode was stencil printed with Ag|AgCl ink. Then, Prussian blue was formed on the working electrode either by drop casting or by electrodeposition, and glucose oxidase was drop cast on top. At this point, the microfabrication process was resumed, and the microfluidics were printed on top of the modified electrodes to complete the construction of hybrid electrochemical fluidic fused filament fabricated devices (h-eF4Ds). This print-pause-print approach is not limited to ink-based electrodes or glucose oxidase, and we envisage these results will pave the way for the effective integration of electrodes in microfluidic devices in a simple and clean-room-free approach, allowing the development of highly customized eF4Ds for a plethora of analytes with high significance.