Fabrication Protocol Research Articles

Cost-Effective Smart Window: Transparency Modulation via Surface Contact Angle Controlled Mist Formation

Implementing simple and inexpensive energy-saving smart technologies in households is quite effective to accomplish on-demand privacy control and reduction in energy consumption. Conventional smart glasses face difficulty in making inroads into the consumer market due to utilizing expensive active layers, electrolytes, and transparent electrodes. Thus, the need of the hour is to develop an unconventional smart window, which should be cost-effective, power-efficient, and simple to fabricate. Against this backdrop, we report the fabrication of a new class of smart partition windows termed "mist-driven transparency switching glass". The fabrication protocol includes surface energy modification of two glass panes, followed by assembling them into a square or rectangular-shaped narrow cell with appropriate inlets and outlets for mist. In its pristine state, the device is transparent, as expected of two plain glasses forming a cell. Insertion of cool mist into the device produces tiny droplets onto the inner walls due to condensation enabling scattering of light, thereby producing the translucent state. The optimized device shows a transmittance modulation of as much as ∼65% at 550 nm, allowing it to reduce the indoor temperature by more than 30% compared to a regular glass windowpane. To realize commercial viability, a large area device (30 × 30 cm2) was fabricated, which could be operated wirelessly through a cellphone application paving the way for incorporating the Internet of Things into the technology.

Thin-film conformal fluorescent SU8-phenylenediamine.

The SU8 polymer is a negative photoresist widely used to produce high-quality coatings, with controllable thicknesses ranging from nanometers to millimeters, depending on fabrication protocols. Apart from conventional use cases in microelectronics and fluidics, SU8 is quite an attractive platform in nanophotonics. This material, being straightforwardly processed by ultraviolet lithography, is transparent to wavelengths longer than 500 nm. However, introducing fluorescent agents within the SU8 matrix remains a challenge owing to its high hydrophobicity. Here, we develop a process, where colorful quantum dots co-participate in the polymerization process by epoxide amination and become a part of a new fluorescent material - SU8-phenylenediamine. Through comprehensive characterization methods, including XPS and 1H-NMR analyses, we demonstrate that m-PD covalently binds to SU8 epoxy sites with its molecular amine, virtually forming a new material and not just a mixture of two compounds. After characterizing the new strongly fluorescent platform, thin 300 nm films were created on several surfaces, including a conformal coverage of a nanofluidic capillary. This new process provides opportunities to incorporate various functional molecules into optoelectronic devices without the need for multistep deposition and surface functionalization.

Encapsulation of MSCs in PRP-Derived Fibrin Microbeads.

Platelet-rich plasma (PRP) is a highly concentrated platelet-containing blood plasma that incorporates a significant amount of growth factors and cytokines needed to accelerate the tissue repair process. PRP has been used effectively for many years in the treatment of various wounds by direct injection into the target tissue or impregnation with scaffold or graft materials. Since autologous PRP can be obtained by simple centrifugation, it is an attractive and inexpensive product for use in repairing damaged soft tissues. Cell-based regenerative approaches, which draw attention in the treatment of tissue and organ injuries, are based on the principle of delivering stem cells to damaged sites by various means, including encapsulation. Current biopolymers used in cell encapsulation have some advantages with some limitations. By adjusting its physicochemical properties, PRP-derived fibrin can become an efficient matrix material for encapsulating stem cells. This chapter covers the fabrication protocol of PRP-derived fibrin microbeads and their use to encapsulate stem cells as a general bioengineering platform for prospective regenerative medical applications.

Simultaneous detection of neurotransmitters and Cu2+ using double-bore carbon fiber microelectrodes via fast-scan cyclic voltammetry.

There is a great demand to broaden our understanding of the multifactorial complex etiology of neurodegenerative diseases to aid the development of more efficient therapeutics and slow down the progression of neuronal cell death. The role of co-transmission and the effect of environmental factors on such diseases have yet to be explored adequately, mainly due to the lack of a proper analytical tool that can perform simultaneous multi-analyte detection in real time with excellent analytical parameters. In this study, we report a simple fabrication protocol of a double-bore carbon-fiber microelectrode (CFM) capable of performing rapid simultaneous detection of neurotransmitters and Cu2+via fast-scan cyclic voltammetry (FSCV) in Tris buffer. After imaging our CFMs via optical microscopy and scanning electron microscopy to ensure the intact nature of the two electrodes in our electrode composite, we performed a detailed analysis of the performance characteristics of our double-bore CFM in five different analyte mixtures, Cu2+-5HT, Cu2+-DA, Cu2+-AA, 5-HT-DA, and 5-HT-AA in Tris buffer, by applying different analyte-specific FSCV waveforms simultaneously. Calibration curves for each analyte in each mixture were plotted while extracting the analytical parameters such as the limit of detection (LOD), linear range, and sensitivity. We also carried out a control experiment series for the same mixtures with single-bore CFMs by applying one waveform at a time to compare the capabilities of our double-bore CFMs. Interestingly, except for the Cu2+-DA solution, all other combinations showed improved LOD, linear ranges, and sensitivity when detecting simultaneously with double-bore CFMs compared to single-bore CFMs, an excellent finding for developing this sensor for future in vivo applications.

Fabrication of curcumin-loaded magnetic PEGylated-PLGA nanocarriers tagged with GRGDS peptide for improving anticancer activity

Carrier-mediated drug delivery systems are highly promising as a treatment option for the targeted delivery of potent cytotoxic drugs with increased efficacy and safety. Considering that poly (lactic-co-glycolic acid) (PLGA) and polyethylene glycol (PEG) polymers each provide certain advantages for biological purposes, PEGylated-PLGA nanoparticles have emerged as a leading candidate among other alternatives. Furthermore, these nanoparticles can be modified with the specific short peptide sequences such as glycine-arginine-glycine-aspartic acid‑serine (GRGDS), which selectively binds to integrins overexpressed in most cancer cells, allowing for targeted delivery. Here, we reported the details in fabrication and characterization of magnetic PEGylated-PLGA nanoparticles functionalized with GRGDS peptide. In addition, superparamagnetic iron oxide nanoparticles (SPIONs) and the natural pharmaceutical compound curcumin (Cur) were loaded into these polymeric nanoparticles to assess their anticancer activity potential. Overall, this study provides comprehensive methodologies, including all synthesis procedures, challenges, and useful suggestions for peptide-conjugated polymeric nanoparticles that may be used for cellular targeting and therapeutic applications.•Step by step fabrication protocol for the Cur loaded magnetic PEGylated-PLGA nanoparticles was presented.•Validation of the fabrication and the GRGDS conjugation to the nanoparticles were shown via detailed characterization studies.•The cytotoxic effect of the Cur-loaded and GRGDS-tagged magnetic nanoparticles was tested on T98G glioblastoma cell line as a preliminary in vitro study.

Membrane-based purification and recovery of phosphate and antibiotics by two-dimensional zeolitic nanoflakes.

The pervasive presence of persistent contaminants in water resources, including phosphate and antibiotics, has attracted significant attention due to their potential adverse effects on ecosystems and human health. Adsorption membranes packed with metal-organic frameworks (MOFs) have been proposed as a potential solution to this challenge due to their high surface area to volume ratio, and the tailored functionality they can provide for selective purification. However, devising a straightforward method to enhance the stability of MOF membranes on polymer supports and manipulate their surface morphology remains challenging. In this study, we present a facile solution immersion technique to fabricate a ZIF-L adsorption membrane on commercial supports by leveraging the self-polymerization characteristics of dopamine. The simple coating methodology provides a polydopamine-lined interface that regulates the ZIF-L heteroepitaxial growth, along with tailored nanoflake morphology. Compared with crystals prepared in bulk solution, the sorbents grown on the membrane exhibit a higher saturation capacity of 248 mg g-1 of phosphate (∼80 mg phosphorus per g sorbent) and 196 mg g-1 for tetracycline in static adsorption experiments at 30 °C. Additionally, the membranes are capable of selectively removing 99.5% of the phosphate in simulant solutions comprising competitive background ions in various concentrations, and efficiently removing tetracycline. The result from the static adsorption experiments directly translates to a flow-through process, showcasing the utility of a composite membrane with a 3 μm thick active layer in practical adsorption applications. The facile solution immersion fabrication protocol introduced in this work may offer a more efficient paradigm to harness the potential of MOF composite membranes in selective adsorption and resource recovery applications.

Shape memory polymers in osteochondral tissue engineering

Bone and cartilage injuries are significantly increasing with population aging. Tissue engineering is considered an alternative and promising approach for alleviating osteochondral tissue injuries along with available therapeutic modalities. 3D- and 4D-printing fabrication protocols have been used to facilitate the production of bone/cartilage scaffolds that are similar to bone and cartilage microenvironments. In this regard, advanced biomaterials, including smart polymers and stimuli-responsive polymers are the first essential elements for improved bone/cartilage regeneration. Shape-memory polymers, are stimuli-responsive materials and are available in permanent and temporary structures. The application of shape-memory scaffolds can lead to providing in vivo-like conditions and improve cell bioactivity and phenotype acquisition. In this review article, we tried to highlight stimuli-responsive polymers and their application in osteochondral tissue engineering.

Digital Diagnostic Wax-up as a Part of Esthetic Dental Treatment in Front Area: a case report

Introduction Digital diagnostic wax up is a 3D representation of the final dental treatment result as far as esthetics (shape, size, color, position, outline), occlusion and function, marginal adaptation are concerned and it is overlaid onto a 3D digital pre-op model. Aim To present the role of the digital diagnostic wax up as a part of dental treatment of front teeth based on a case report Materials and Methods Dental treatment in the front area due to esthetic indications is reported. A young patient is seeking for a whiter and brighter smile. Results The clinical protocol for fabrication of digital diagnostic wax up used in this case is described. Although the veneers provide better long term esthetic result, the patient chose bonding due to no prep approach and lower expenses. The digital diagnostic wax-up was used for printing a 3D model which is needed for fabrication of a silicone index for the composite resin injectable technique. Conclusion The digital diagnostic wax up helps the dentist and the patient to visualize the desired changes. The communication between dental office, lab and patient is a lot easier. Editing is done easily and fast using the software until the perfect result is achieved and accepted by the whole team. It is a precise, time and money saving method that leads to predictable final result and satisfaction of the dental treatment.

Nanoarchitectonics for Free‐Standing Polyelectrolyte Multilayers Films: Exploring the Flipped Surfaces

AbstractThe competency of polyelectrolyte multilayer films (PEM) self‐assembly to modulate the surface architecture at the molecular level has emerged as a captivating technique for various biomedical applications. PEM advancement in bio‐applications covered the development of bio‐sensors, stimuli‐responsive drug‐delivery systems, surface modification for creating antibacterial surfaces, and tissue‐engineering architectures. Different PEM blends, from natural to synthetic ones, have been explored to make a free‐standing version of the PEM stack. This review provides a deeper understanding of the free‐standing PEM (FS‐PEM), their fabrication strategies, and essential properties for specified bio‐applications. The most crucial aspect of fabricating FS‐PEM is choosing an application‐specific fabrication protocol to delaminate the PEM stack from the underlying substrates. Post‐fabrication, there are important considerations to improve the mechanical stability of these FS‐PEMs by crosslinking or adding desired nanoparticles. Overall, essential insights are discussed here for using FS‐PEM for specific bio‐application, which has received little attention in the material science community until now.

Protein-stabilized bimetallic Au/Ag nanoclusters as fluorescent reporters: Synthesis, characterization and their interactions with biocolloids

Fabrication protocol of bovine serum albumin-(BSA) and lysozyme-(LYZ) stabilized gold/silver bimetallic nanoclusters (Au/Ag NCs) have been reported via optimization of the protein/metal ion ratios, metal concentration, pH and incubation time. No other reducing and stabilizing compounds were used to produce these nanoclusters at room temperature. Incorporation of small quantities of silver (∼3–10 w%) into the gold core substantially changed the optical features such as the enhancing of the quantum yield and blue shifting of the emission wavelength compared to their purely gold-based derivatives. Applicability of the BSA- and LYZ-directed Au/Ag NCs having intense fluorescence at above ∼ 600 nm as potential fluorescent reporters (FRs) has been studied by examining their interactions with chitosan/hyaluronic acid (Chit/HyA)-based colloidal drug delivery particles. It was found that, in contrast to the monometallic derivatives, our bimetallic NCs, prepared at room temperature and stabilized with larger amounts of proteins, exhibit a much higher degree of stability during interaction with biocolloids. The results are confirmed by fluorescence measurements in addition to light scattering studies, since the aggregation-induced emission is not observed dominantly, as clearly confirmed for Au-based NCs. Utilizing the alloying process and the milder preparation conditions can be a promising route to the synthesis of noble metal NCs-based FRs in biomedical field.

Fused optical fiber combiner based on indium fluoride glass: perspectives for mid-IR applications.

For the first time, to the best of our knowledge, the design and characterization of a 3 × 1 fused fiber combiner based on multimode step-index fluoroindate optical fibers (InF3) has been performed. Several efforts to develop a well-consolidated normalization procedure and a fabrication protocol have been required due to the low melting temperature and the mechanical properties of fluoroindates. Fabrication results demonstrate repeatability and absence of crystallization. Therefore, the described fabrication process paves the way for manufacturing fluoroindate devices. The electromagnetic design of the combiner is carried out through modal investigation and beam propagation method by computing the transmission efficiency. The experimental results agree with the simulation and demonstrate the device feasibility to operate in the mid-infrared spectral range.

An open-source LED array illumination system for automated multiwell plate cell culture photodynamic therapy experiments

Photodynamic therapy (PDT) research would benefit from an automated, low-cost, and easy-to-use cell culture light treatment setup capable of illuminating multiple well replicates within standard multiwell plate formats. We present an LED-array suitable for performing high-throughput cell culture PDT experiments. The setup features a water-cooling loop to keep the LED-array temperature nearly constant, thus stabilizing the output power and spectrum. The setup also features two custom-made actuator arms, in combination with a pulse-width-modulation (PWM) technique, to achieve programmable and automatic light exposures for PDT. The setup operates at ~ 690 nm (676–702 nm, spectral output full-width half-maximum) and the array module can be readily adapted to other LED wavelengths. This system provides an illumination field with adjustable irradiance up to 400 mW/cm2 with relatively high spectral and power stability comparing with previously reported LED-based setups. The light doses provided by the LED array were validated with comparison to traditional laser PDT. This open-source illumination platform (including the detailed technical description, fabrication protocols, and parts list provided here) helps to make custom light sources more accessible and of practical use for photomedicine research.

Optimization of 3D-aggregated spheroid model (3D-ASM) for selecting high efficacy drugs

Various three-dimensional (3D) cell culture methods have been developed to implement tumor models similar to in vivo. However, the conventional 3D cell culture method has limitations such as difficulty in using an extracellular matrix (ECM), low experimental reproducibility, complex 3D cell culture protocol, and difficulty in applying to high array plates such as 96- or 384-plates. Therefore, detailed protocols related to robust 3D-aggregated spheroid model (3D-ASM) production were optimized and proposed. A specially designed wet chamber was used to implement 3D-ASM using the hepatocellular carcinoma (HCC) cell lines, and the conditions were established for the icing step to aggregate the cells in one place and optimized ECM gelation step. Immunofluorescence (IF) staining is mainly used to simultaneously analyze drug efficacy and changes in drug-target biomarkers. By applying the IF staining method to the 3D-ASM model, confocal microscopy imaging and 3D deconvolution image analysis were also successfully performed. Through a comparative study of drug response with conventional 2D-high throughput screening (HTS), the 3D-HTS showed a more comprehensive range of drug efficacy analyses for HCC cell lines and enabled selective drug efficacy analysis for the FDA-approved drug sorafenib. This suggests that increased drug resistance under 3D-HTS conditions does not reduce the analytical discrimination of drug efficacy, also drug efficacy can be analyzed more selectively compared to the conventional 2D-HTS assay. Therefore, the 3D-HTS-based drug efficacy analysis method using an automated 3D-cell spotter/scanner, 384-pillar plate/wet chamber, and the proposed 3D-ASM fabrication protocol is a very suitable platform for analyzing target drug efficacy in HCC cells.

Block Copolymer-based Photonic Pigments: Towards Structural Non-iridescent Brilliant Coloration

Creating color through the self-assembly of specific building blocks to fabricate photonic morphologies is a promising and intriguing approach to reproducing the flamboyant visual effects and dynamic properties observed in the natural world. However, the complexity and lack of robustness in the manufacture of these nanostructured materials hinder their technical exploitation on a large scale. To overcome such limitations, here we present a novel methodology to create bioinspired photonic pigments as dispersed and micrometer-scale particles based on highly ordered concentric lamellar microspheres made of block copolymers. First, we introduce the fabrication protocol and the advantages of this approach compared to the traditional colloidal self-assembly. Then, we discuss some possible future research directions focused on developing hybrid organic-inorganic photonic pigments with enhanced dielectric contrast, reduced scattering, and specific functionalities. Finally, we speculate on possible applications for these structures that go beyond their use as simple photonic pigments.

Optimization Strategies for Commercialization of High-Temperature Pemfcs

The increasing requirement for renewable energy places high-temperature proton exchange membrane fuel cells (HT-PEMFCs) on the forefront of “green” energy-generating power devices. Compared to certain PEMFC technologies, HT-PEMFCs possess faster electrode kinetics, high tolerance to fuel poisons and impurities, no humidification requirements, simplified cooling and system design.1 Herein we present optimization strategies of membrane electrode assemblies (MEAs) for HT-PEMFCs, focusing mainly on the (1) component characterization, (2) MEA fabrication, and (3) testing protocols. A selection of gas diffusion electrodes (GDEs) was tested, as well as the presence/absence of a microporous layer (MPL) on the cell performance. The catalyst layer (CL) has been modified in the terms of fabrication methods, catalyst type, platinum (Pt) content, as well as a variety of binders and additives. The PEM optimizations focused on varying and testing: (1) the molecular weight (Mw) of polybenzimidazole (PBI) for membrane casting, (2) PEM thickness, and its (3) acid doping levels (ADL). The MEAs were then fabricated at altering hot-pressing conditions and their performances were compared. The MEA testing focused on activation, humidity levels, high current operations, elevated temperature and pressure operations, as well as long-term stationary and dynamic durability protocol studies. The material fabrication optimizations resulted in higher onset and peak-of-life voltages, as well as longer cell durability. Together with optimized cell testing, we have achieved power densities of more than 0.8 W/cm2, with stationary durability of more than 13,000 hours at 0.3 A/cm2 current density, with the degradation rate of 4 μV/h. Originally the dynamic durability was demonstrated with more than 290 start-stop cycles showing the performance degradation of up to 100 µV/cycle, however, new tests show degradation rates of less than 46 µV/cycle. The performance and durability that we have demonstrated here position HT-PEMFCs as matured technology that can enter the mass market as a commercial and reliable electrochemical power device, to answer the ever-increasing societal energy demands in the age of climate change. Figure 1. Performance of Blue World Technologies MEAs: (A) from different production batches showing small performance scatter, and (B) i-V and power curves at elevated pressures.Reference:1. O. Jensen, D. Aili, H. A. Hjuler, Q. Li, High Temperature Polymer Electrolyte Membrane Fuel Cells - Approaches, Status and Perspective, ISBN 978-3-319-17081-7; DOI 10.1007/978-3-319-17082-4, Springer International Publishing, New York, 2015. Figure 1

Polymeric Microfluidic Devices Fabricated Using Epoxy Resin for Chemically Demanding and Day-Long Experiments.

Polydimethylsiloxane (PDMS) is a widely used material in laboratories for fabricating microfluidic devices with a rapid and reproducible prototypingability, owing to its inherent properties (e.g., flexibility, air permeability, and transparency). However, the PDMS channel is easily deformed under pressures applied to generate flows because of its elasticity, which can affect the robustness of experiments. In addition, air permeability of PDMS causes the pervaporation of water, and its porous structure absorbs oil and even small hydrophobic molecules, rendering it inappropriate for chemically demanding or day-long experiments. In this study, we develop a rapid and reproducible fabrication method for polymer-based rigid microfluidic devices, using epoxy resin that can overcome the limitations of PDMS channels, which are structurally and chemically robust. We first optimize a high-resolution fabrication protocol to achieve convenient and repeatable prototyping of polymeric devices via epoxy casting using PDMS soft molds. In addition, we compare the velocity changes in PDMS microchannels by tracking fluorescent particles in various flows (~133 μL/min) to demonstrate the structural robustness of the polymeric device. Furthermore, by comparing the adsorption of fluorescent hydrophobic chemicals and the pervaporation through channel walls, we demonstrate the excellent chemical resistance of the polymeric device and its suitability for day-long experiments. The rigid polymeric device can facilitate lab-on-chip research and enable various applications, such as high-performance liquid chromatography, anaerobic bacterial culture, and polymerase chain reaction, which require chemically or physically demanding experiments.

Multiplexed Electrochemical Device for the Detection of Biomarkers of Parkinson’s Disease Using 3D Printing

Acetylcholine and dopamine are two important neurotransmitters when diagnosing Parkinson’s disease, as the imbalance in acetylcholine and dopamine levels can lead to progressive neurological disorders. Currently, there are many detection methods for the individual neurotransmitters with their own advantages and disadvantages. Previously, dopamine has been highlighted as the most relevant biomarker for diagnosis, but interest in combined detection of acetylcholine and dopamine has increased. Since it has been discovered that both motor function and learning difficulties in Parkinsonism is dependent on dopamine and acetylcholine levels, determination of both biomarkers simultaneously has increased therapeutic relevance. This work reports the detection of acetylcholine and dopamine using 3D printed potentiometric and 3D printed voltammetric sensors, respectively. Linear responses were obtained over broad concentration ranges for both biomarkers with a fully 3D printed ion-selective membrane (acetylcholine) and conductive carbon electrodes (dopamine). This work describes a novel fabrication protocol for sensors that have the capability to detect both biomarkers simultaneously when integrated into a low-cost, reliable, and user-friendly multiplexed device, also fabricated with 3D printing technologies.

Direct Comparison of the Effect of Processing Conditions in Electrolyte-Gated and Bottom-Gated TIPS-Pentacene Transistors

Among the plethora of soluble and easy processable organic semiconductors, 6,13-Bis(triisopropylsilylethynyl)pentacene (TIPS-P5) is one of the most promising materials for next-generation flexible electronics. However, based on the information reported in the literature, it is difficult to exploit in field-effect transistors the high-performance characteristics of this material. This article correlates the HMDS functionalization of the silicon substrate with the electrical characteristics of TIPS-P5-based bottom gate organic field-effect transistors (OFETs) and electrolyte-gated organic field-effect transistors (EGOFETs) fabricated over the same platform. TIPS-P5 transistors with a double-gate architecture were fabricated by simple drop-casting on Si/SiO2 substrates, and the substrates were either functionalized with hexamethyldisilazane (HMDS) or left untreated. The same devices were characterized both as standard bottom-gate transistors and as (top-gate) electrolyte-gated transistors, and the results with and without HMDS treatment were compared. It is shown that the functionalization of the silicon substrate negatively influences EGOFETs performance, while it is beneficial for bottom-gate OFETs. Different device architectures (e.g., bottom-gate vs. top-gate) require specific evaluation of the fabrication protocols starting from the effect of the HMDS functionalization to maximize the electrical characteristics of TIPS-P5-based devices.

High-Throughput Nanopore Fabrication and Classification Using Xe-Ion Irradiation and Automated Pore-Edge Analysis.

Large-area nanopore drilling is a major bottleneck in state-of-the-art nanoporous 2D membrane fabrication protocols. In addition, high-quality structural and statistical descriptions of as-fabricated porous membranes are key to predicting the corresponding membrane-wide permeation properties. In this work, we investigate Xe-ion focused ion beam as a tool for scalable, large-area nanopore fabrication on atomically thin, free-standing molybdenum disulfide. The presented irradiation protocol enables designing ultrathin membranes with tunable porosity and pore dimensions, along with spatial uniformity across large-area substrates. Fabricated nanoporous membranes are then characterized using scanning transmission electron microscopy imaging, and the observed nanopore geometries are analyzed through a pore-edge detection and analysis script. We further demonstrate that the obtained structural and statistical data can be readily passed on to computational and analytical tools to predict the permeation properties at both individual pore and membrane-wide scales. As an example, membranes featuring angstrom-scale pores are investigated in terms of their emerging water and ion flow properties through extensive all-atom molecular dynamics simulations. We believe that the combination of experimental and analytical approaches presented here will yield accurate physics-based property estimates and thus potentially enable a true function-by-design approach to fabrication for applications such as osmotic power generation and desalination/filtration.

A Protocol for Fabrication and on-Chip Cell Culture to Recreate PAH-Afflicted Pulmonary Artery on a Microfluidic Device.

Pulmonary arterial hypertension (PAH) is a rare pulmonary vascular disease that affects people of all ethnic origins and age groups including newborns. In PAH, pulmonary arteries and arterioles undergo a series of pathological changes including remodeling of the entire pulmonary vasculatures and extracellular matrices, mis-localized growth of pulmonary arterial cells, and development of glomeruloid-like lesions called plexiform lesions. Traditionally, various animal and cellular models have been used to understand PAH pathophysiology, investigate sex-disparity in PAH and monitor therapeutic efficacy of PAH medications. However, traditional models can only partially capture various pathological features of PAH, and they are not adaptable to combinatorial study design for deciphering intricately intertwined complex cellular processes implicated in PAH pathogenesis. While many microfluidic chip-based models are currently available for major diseases, no such disease-on-a-device model is available for PAH, an under investigated disease. In the absence of any chip-based models of PAH, we recently proposed a five-channel polydimethylsiloxane (PDMS)-based microfluidic device that can emulate major pathological features of PAH. However, our proposed model can make a bigger impact on the PAH field only when the larger scientific community engaged in PAH research can fabricate the device and develop the model in their laboratory settings. With this goal in mind, in this study, we have described the detailed methodologies for fabrication and development of the PAH chip model including a thorough explanation of scientific principles for various steps for chip fabrication, a detailed list of reagents, tools and equipment along with their source and catalogue numbers, description of laboratory setup, and cautionary notes. Finally, we explained the methodologies for on-chip cell seeding and application of this model for studying PAH pathophysiology. We believe investigators with little or no training in microfluidic chip fabrication can fabricate this eminently novel PAH-on-a-chip model. As such, this study will have a far-reaching impact on understanding PAH pathophysiology, unravelling the biological mystery associated with sexual dimorphism in PAH, and developing PAH therapy based on patient sex and age.