PDMS Device Research Articles

A miniaturized multicellular platform to mimic the 3D structure of the alveolar-capillary barrier.

Several diseases affect the alveoli, and the efficacy of medical treatments and pharmaceutical therapies is hampered by the lack of pre-clinical models able to recreate in vitro the diseases. Microfluidic devices, mimicking the key structural and compositional features of the alveoli, offer several advantages to medium and high-throughput analysis of new candidate therapies. Here, we developed an alveolus-on-a-chip recapitulating the microanatomy of the physiological tissue by including the epithelium, the fibrous interstitial layer and the capillary endothelium. A PDMS device was obtained assembling a top layer and a bottom layer obtained by replica molding. A polycaprolactone/gelatin (PCL-Gel) electrospun membrane was included within the two layers supporting the seeding of 3cell phenotypes. Epithelial cells were grown on a fibroblast-laden collagen hydrogel located on the top side of the PCL-Gel mats while endothelial cells were seeded on the basolateral side of the membrane. The innovative design of the microfluidic device allows to replicate both cell-cell and cell-extracellular matrix interactions according to the in vivo cell arrangement along with the establishment of physiologically relevant air-liquid interface conditions. Indeed, high cell viability was confirmed for up to 10 days and the formation of a tight endothelial and epithelial barrier was assessed by immunofluorescence assays.

New design of microchip for in-situ DLS measurement of colloidal suspensions

Stability of dispersed materials remains an important question in a wide variety of fields such as cosmetics, catalysis, food or energy and the environment. As stability is directly linked to the size of the dispersed colloids it is essential to assess the size distribution of colloidal suspensions. Nowadays, microfluidic-based approaches generate increasing interest as they represent flexible and fast measurements allowing high throughput experimentations. However, characterization of colloidal dispersions is usually performed by dynamic light scattering (DLS), that requires static measurements as well as significant volumes, that are not compatible with on-line analysis and microfluidics. Moreover, due to flow-induced decorrelation terms, DLS measurements in microfluidic channels are only accurate at very low shear rates. This work aimed at developing an on-line microfluidic device for dispersed materials characterization using DLS. The main challenges of this project were i) to adapt the microfabrication of the PDMS device in order to combine microchannels of hundreds of microns with a milli-fluidic cavity to perform the DLS measurements, and ii) to downsize the DLS set up. A PDMS microchip, consisting in a millimeter cavity for DLS measurements in parallel with a microchannel, was designed to perform the measurement on the sample without stopping the suspension flow during the microfluidic experiment. The cavity geometry was then optimized thanks to numerical simulation to ensure a good sweep efficiency and to downscale as much as possible without impairing the DLS signal. By adapting the microfabrication process, a PDMS microfluidic chip was designed allowing the size measurements of successive suspensions containing 100 and 12 ​nm diameter particles. This work is a first step towards the implementation of a new technological building block for online microfluidic characterization. © 2017 Elsevier Inc. All rights reserved.

A total-internal-reflection-based Fabry–Pérot resonator for ultra-sensitive wideband ultrasound and photoacoustic applications

In photoacoustic and ultrasound imaging, optical transducers offer a unique potential to provide higher responsivity, wider bandwidths, and greatly reduced electrical and acoustic impedance mismatch when compared with piezoelectric transducers. In this paper, we propose a total-internal-reflection-based Fabry–Pérot resonator composed of a 12-nm-thick gold layer and a dielectric resonant cavity. The resonator uses the same Kretschmann configuration as surface plasmon resonators (SPR). The resonators were analyzed both theoretically and experimentally. The experimental results were compared with those for an SPR for benchmarking. The 1.9-μm-thick-PMMA- and 3.4-μm-thick-PDMS-based resonators demonstrated responsivities of 3.6- and 30-fold improvements compared with the SPR, respectively. The measured bandwidths for the PMMA, PDMS devices are 110 MHz and 75 MHz, respectively. Single-shot sensitivity of 160 Pa is obtained for the PDMS device. The results indicate that, with the proposed resonator in imaging applications, sensitivity and the signal-to-noise ratio can be improved significantly without compromising the bandwidth.

Bi-directional Dual-flow-RootChip for Physiological Analysis of Plant Primary Roots Under Asymmetric Perfusion of Stress Treatments.

Due to technical limitations, research to date has mainly focused on the role of abiotic and biotic stress-signalling molecules in the aerial organs of plants, including the whole shoot, stem, and leaves. Novel experimental platforms including the dual-flow-RootChip (dfRC), PlantChip, and RootArray have since expanded this to plant-root cell analysis. Based on microfluidic platforms for flow stream shaping and force sensing on tip-growing organisms, the dfRC has further been expanded into a bi-directional dual-flow-RootChip (bi-dfRC), incorporating a second adjacent pair of inlets/outlet, enabling bi-directional asymmetric perfusion of treatments towards plant roots (shoot-to-root or root-to-shoot). This protocol outlines, in detail, the design and use of the bi-dfRC platform. Plant culture on chip is combined with guided root growth and controlled exposure of the primary root to solute changes. The impact of surface treatment on root growth and defence signals can be tracked in response to abiotic and biotic stress or the combinatory effect of both. In particular, this protocol highlights the ability of the platform to culture a variety of plants, such as Arabidopsis thaliana, Nicotiana benthamiana, and Solanum lycopersicum, on chip. It demonstrates that by simply altering the dimensions of the bi-dfRC, a broad application basis to study desired plant species with varying primary root sizes under microfluidics is achieved. Key features Expansion of the method developed by Stanley et al. (2018a) to study the directionality of defence signals responding to localised treatments. Description of a microfluidic platform allowing culture of plants with primary roots up to 40 mm length, 550μm width, and 500 μm height. Treatment with polyvinylpyrrolidone (PVP) to permanently retain the hydrophilicity of partially hydrophobic bi-dfRC microchannels, enabling use with surface-sensitive plant lines. Description of novel tubing array setup equipped with rotatable valves for switching treatment reagent and orientation, while live-imaging on the bi-dfRC. Graphical overview Graphical overview of bi-dfRC fabrication, plantlet culture, and setup for root physiological analysis.(a) Schematic diagram depicting photolithography and replica molding, to produce a PDMS device. (b) Schematic diagram depicting seed culture off chip, followed by sub-culture of 4-day-old plantlets on chip. (c) Schematic diagram depicting microscopy and imaging setup, equipped with a media delivery system for asymmetric treatment introduction into the bi-dfRC microchannel root physiological analysis under varying conditions.

Microfluidic dual picoinjection based encapsulation of hemoglobin in alginate microcapsules reinforced by a poly(L-lysine)-g-poly(ethylene glycol).

Hemoglobin (Hb) encapsulation inside polysaccharide hydrogels has been considered a possible red blood cell (RBC) surrogate in transfusiology. Here we report on the microfluidic dual picoinjection assisted synthesis of Hb encapsulated alginate-poly(L-lysine)-g-poly(ethylene glycol) beads. This process is realized by the on-chip injections of blended Hb alginate solutions in emulsified aqueous calcium chloride (CaCl2) droplets followed by a subsequent injection of an aqueous PLL-g-PEG into each emulsified aqueous droplet. The proposed fabrication approach was realized using a flow-focusing and two picoinjection sites in a single PDMS device. Aqueous CaCl2 solution was emulsified and infused with Hb-alginate solution as the squeezed droplet passed through the first picoinjection site. The injection of PLL-g-PEG to reinforce the microgel and minimize the protein leaching was realized in the second picoinjection site located downstream from the first in the same microfluidic channel. In this process, monodisperse Hb-alginate-PLL-g-PEG particles with a diameter around the size of RBCs (9 μm) were obtained with around 80% of the 7.5 mg ml-1 Hb included in the injected aqueous alginate retaining in the obtained microparticles. Microparticles with Hb loading (32.8 pg per bead) and retention (28.8 pg per bead) over a week of storage at 4 °C are in accordance with the average amount of Hb per RBC. The Hb-alginate-PLL-g-PEG microbeads fabricated in the size range of RBCs are significant for further exploration.

Fabrication of Wearable PDMS Device for Rapid Detection of Nucleic Acids via Recombinase Polymerase Amplification Operated by Human Body Heat.

Pathogen detection by nucleic acid amplification proved its significance during the current coronavirus disease 2019 (COVID-19) pandemic. The emergence of recombinase polymerase amplification (RPA) has enabled nucleic acid amplification in limited-resource conditions owing to the low operating temperatures around the human body. In this study, we fabricated a wearable RPA microdevice using poly(dimethylsiloxane) (PDMS), which can form soft—but tight—contact with human skin without external support during the body-heat-based reaction process. In particular, the curing agent ratio of PDMS was tuned to improve the flexibility and adhesion of the device for better contact with human skin, as well as to temporally bond the microdevice without requiring further surface modification steps. For PDMS characterization, water contact angle measurements and tests for flexibility, stretchability, bond strength, comfortability, and bendability were conducted to confirm the surface properties of the different mixing ratios of PDMS. By using human body heat, the wearable RPA microdevices were successfully applied to amplify 210 bp from Escherichia coli O157:H7 (E. coli O157:H7) and 203 bp from the DNA plasmid SARS-CoV-2 within 23 min. The limit of detection (LOD) was approximately 500 pg/reaction for genomic DNA template (E. coli O157:H7), and 600 fg/reaction for plasmid DNA template (SARS-CoV-2), based on gel electrophoresis. The wearable RPA microdevice could have a high impact on DNA amplification in instrument-free and resource-limited settings.

Photolithography-free fabrication of photoresist-mold for rapid prototyping of microfluidic PDMS devices

Traditional soft lithography based PDMS device fabrication requires complex procedures carried out in a clean room. Herein, we report a photolithography-free method that rapidly produces PDMS devices in 30 min. By using a laser cutter to ablate a tape, a male photoresist mold can be obtained within 5 min by a simple heating-step, which offers significant superiority over currently used photolithography-based method. Since it requires minimal energy to cut the tape, our fabrication strategy shows good resolution (∼ 100 µm) and high throughput. Furthermore, the micro-mold height can be easily controlled by changing the tape types and layers. As a proof-of-concept, we demonstrated that the fabricated PDMS devices are compatible with biochemical reactions such as quenching reaction of KI to fluorescein and cell culture/staining. Collectively, our strategy shows advantages of low input, simple operation procedure and short fabrication time, therefore we believe this photolithography-free method could serve as a promising way for rapid prototyping of PDMS devices and be widely used in general biochemical laboratories.

Nanoadhesive layer to prevent protein absorption in a poly(dimethylsiloxane) microfluidic device.

Poly(dimethylsiloxane) (PDMS) is widely used as a microfluidics platform material; however, it absorbs various molecules, perturbing specific chemical concentrations in microfluidic channels. We present a simple solution to prevent adsorption into a PDMS microfluidic device. We used a vapor-phase-deposited nanoadhesive layer to seal PDMS microfluidic channels. Absorption of fluorescent molecules into PDMS was efficiently prevented in the nanolayer-treated PDMS device. Importantly, when cultured in a nanolayer-treated PDMS device, yeast cells exhibited the expected concentration-dependent response to a mating pheromone, including mating-specific morphological and gene expression changes, while yeast cultured in an untreated PDMS device did not properly respond to the pheromone. Our method greatly expands microfluidic applications that require precise control of molecule concentrations.

Oxygen Monitor to Study Vascularization of Medical Devices

Prevascularized medical devices can improve cell therapy. Such devices may replace whole organ transplantation with hosting only the necessary therapeutic cells. We have developed a noninvasive optical technology to study the vascularization into such medical devices. In our technique, oxygen partial pressure within a device is monitored by Oxygen Sensitive Tubes (OSTs), comprising oxygen permeable silicone tubing with inner luminal surfaces coated by an oxygen-sensitive porphyrin dye. OSTs were placed within a PDMS device and transplanted into the subcutaneous space of athymic nude mice. An optical probe placed over the skin excites the OSTs with a pulse of light and detects the luminescent lifetime of emitted light, which is uniquely related to oxygen partial pressure. Furthermore, we developed a Dynamic Inhalation Gas Test (DIGT) to determine the oxygen transport rate between the microvasculature and the device. DIGT works by monitoring oxygen partial pressure in a device following a step change in inhaled-gas oxygen content. We report DIGT oxygen dynamics measured intermittently over eight weeks. Our study shows DIGT dynamics are unique to each implant, supporting the important role of the host tissue response in the availability of oxygen over time.

3D-printed microfluidic manipulation device integrated with magnetic array

This paper reported a transparent, high-precision 3D-printed microfluidic device integrated with magnet array for magnetic manipulation. A reserved groove in the device can well constrain the Halbach array or conventional alternating array. Numerical simulations and experimental data indicate that the magnetic flux density ranges from 30 to 400 mT and its gradient is about 0.2–0.4 T/m in the manipulation channel. The magnetic field parameters of Halbach array in the same location are better than the other array. Diamagnetic polystyrene beads experience a repulsive force and move away from the magnetic field source under the effect of negative magnetophoresis. It is undeniable that as the flow rate increases, the ability of Halbach array to screen particle sizes decreases. Even so, it has a good particle size discrimination at a volumetric flow rate of 1.08 mL/h, which is much larger than that of a conventional PDMS device with a single magnet. The observed particle trajectories also confirm these statements. The deflection angle is related to the magnetic field, flow rate, and particle size. This 3D-printed device integrated with Halbach array offers excellent magnetic manipulation performance.

Change of Preferential Liquid Breakthrough Pathways in PEMFC Gas Diffusion Layers: Evidence of Pressure-Induced Dynamic Transport

Polymer Electrolyte Membrane (PEM) fuel cells are considered as promising clean sources for automotive applications. A key challenge to reduce the cost of this technology is to increase the power density by operating PEM fuel cells at high current densities. One strategy is to improve the 3D structure of the gas diffusion layer (GDL) located between the channel and the catalyst layer (CL). At the cathode side, this GDL is used to evacuate the liquid water toward the channel while providing access to the oxygen to the CL [1]. Although innovative GDL structures have been proposed, a better understanding of the liquid water would help to design better GDL. In particular, few works have investigated the liquid water transport after several hours of steady state fuel cell operating conditions although changes in liquid water preferential pathway have been observed [2]. Such changes are not predicted in theory, and a physical explanation of these mechanisms is still required. In this presentation, we investigate ex situ the change of liquid water preferential pathways through PEM fuel cell GDLs. Liquid water breakthough from the GDL to the fuel cell channels is mimicked using a microfluidic PDMS device with embedded GDLs. Liquid water is injected through the GDL to the microchannel, and changes in liquid water pathways are observed after several hours of operations. These changes in liquid water pathway are attributed to the dynamic of droplet eruption in fuel cell microchannel. To validate this assumption, a small capillary network with is built. It is shown that after the liquid water breakthrough, the pressure variation due to the growth of the water droplets in the microchannel enables the liquid water to invade smaller capillaries which at the end can change the preferential path. The experimental observations reported in this work show that the invasion percolation theory can be used to predict the preferential pathway at the onset of the liquid water breakthrough, but pressure-induced dynamic transport [3] due the water droplet eruption in the channel has also to be taken into account to predict the preferential pathway after long time of fuel cell operation.

Pattern separation and completion of distinct axonal inputs transmitted via micro-tunnels between co-cultured hippocampal dentate, CA3, CA1 and entorhinal cortex networks

Objective. Functions ascribed to the hippocampal sub-regions for encoding episodic memories include the separation of activity patterns propagated from the entorhinal cortex (EC) into the dentate gyrus (DG) and pattern completion in CA3 region. Since a direct assessment of these functions is lacking at the level of specific axonal inputs, our goal is to directly measure the separation and completion of distinct axonal inputs in engineered pairs of hippocampal sub-regional circuits. Approach. We co-cultured EC-DG, DG-CA3, CA3-CA1 or CA1-EC neurons in a two-chamber PDMS device over a micro-electrode array (MEA60), inter-connected via distinct axons that grow through the micro-tunnels between the compartments. Taking advantage of the axonal accessibility, we quantified pattern separation and completion of the evoked activity transmitted through the tunnels from source into target well. Since pattern separation can be inferred when inputs are more correlated than outputs, we first compared the correlations among axonal inputs with those of target somata outputs. We then compared, in an analog approach, the distributions of correlation distances between rate patterns of the axonal inputs inside the tunnels with those of the somata outputs evoked in the target well. Finally, in a digital approach, we measured the spatial population distances between binary patterns of the same axonal inputs and somata outputs. Main results. We found the strongest separation of the propagated axonal inputs when EC was axonally connected to DG, with a decline in separation to CA3 and to CA1 for both rate and digital approaches. Furthermore, the digital approach showed stronger pattern completion in CA3, then CA1 and EC. Significance. To the best of our knowledge, these are the first direct measures of pattern separation and completion for axonal transmission to the somata target outputs at the rate and digital population levels in each of four stages of the EC-DG-CA3-CA1 circuit.

PDMS-Based Microfluidic Device for Infrared-Transmission Spectro-Electrochemistry

Abstract Polydimethylsiloxane (PDMS) is a well-known material used in making microfluidic devices through soft lithography. PDMS strongly absorbs infrared (IR) light in the mid infrared region, which can be alleviated by using thin-film PDMS devices. However, IR detection with PDMS devices usually requires an IR-transparent backbone material. Here, we report a microfluidic system using a 160 µm PDMS thin-film device (50-µm path length) without a supporting backbone for IR spectroelectrochemistry (IR-SEC). The PDMS device is assembled with four layers: microchannel-supporting window, microchannel thin film, electrode-embedded thin film, and electrode-supporting window. In this work, mapping of ferrocyanide oxidation was investigated under an IR microscope. An electrochemical reaction, occurring between the working and counter electrodes, was observed clearly. Moreover, the electron-transfer process of ferrocyanide oxidation was investigated from the IR absorbance spectrum. This mapping study demonstrates that the relationship between the electrochemical reactants and products partly agree well with the Nernst equation. Accordingly, PDMS thin-film devices have the capability for studying electrochemical reactions by using IR-SEC measurements with low sample consumption.

Fabricating devices with improved adhesion between PDMS and gold-patterned glass

Gold-patterned glass substrates sealed to a PDMS fluidic layer is an architecture widely employed in microfluidic devices. The plasma bonding method is commonly used but presents the disadvantage of being ineffective on gold surfaces, creating a sealing weakness where solution leakage can be a problem. Frequently, alternative approaches to sealing include additional patterned layers that often require expensive cleanroom time. Here we demonstrate a sealing method involving carboxyl-terminated and amine-terminated silanes appropriate for PDMS-based device fabrication with patterned gold/glass surfaces. This cleanroom-free methodology improves solution leak pressure 37-fold compared to the standard plasma bonding technique on these patterned substrates. This bonding strategy is demonstrated in a proof-of-concept electrochemical cell enclosed in a PDMS chamber. An electrochemical method for preparing a clean and electroanalytically useful gold surface in the PDMS device is detailed.

Droplet trapping and fast acoustic release in a multi-height device with steady-state flow.

We demonstrate a novel multilayer polydimethylsiloxane (PDMS) device for selective storage and release of single emulsion droplets. Drops are captured in a microchannel cavity and can be released on-demand through a triggered surface acoustic wave pulse. The surface acoustic wave (SAW) is excited by a tapered interdigital transducer (TIDT) deposited on a piezoelectric lithium niobate (LiNbO3) substrate and inverts the pressure difference across the cavity trap to push a drop out of the trap and back into the main flow channel. Droplet capture and release does not require a flow rate change, flow interruption, flow inversion or valve action and can be achieved in as fast as 20 ms. This allows both on-demand droplet capture for analysis and monitoring over arbitrary time scales, and continuous device operation with a high droplet rate of 620 drops per s. We hence decouple long-term droplet interrogation from other operations on the chip. This will ease integration with other microfluidic droplet operations and functional components.

Hydrodynamic injection on electrophoresis microchips using an electronic micropipette

Here we report for the first time the use of an electronic micropipette as hydrodynamic (HD) injector for microchip electrophoresis (ME) devices. The micropipette was directly coupled to a PDMS device, which had been fabricated in a simple cross format with two auxiliary channels for sample volume splitting. Sample flow during the injection procedure was controlled in automatic dispenser mode using a volume of 0.6µL. Channel width and device configuration were optimized and the best results were achieved using a simple cross layout containing two auxiliary channels with 300µm width for sample splitting. The performance of the HD injector was evaluated using a model mixture of high-mobility cationic species. The results obtained were compared to the data obtained via electrokinetic (EK) injection. Overall, the HD provided better analytical performance in terms of resolution and injection-to-injection repeatability. The relative standard deviation (RSD) values for peak intensities were lower than 5% (n=10) when the micropipette was employed. In comparison with EK injection, the use of the proposed HD injector revealed an unbiased profile for a mixture containing K+ and Li+(300 µmol L−1 each) over various buffer concentrations. For EK injection, the peak areas decreased from 2.92 ± 0.20–0.72 ± 0.14Vs for K+ and from 1.30 ± 0.10–0.38 ± 0.10Vs for Li+ when the running buffer increased from 20 to 50mmolL−1. For HD injection, the peak areas for K+ and Li+ exhibited average values of 2.48±0.07 and 2.10±0.06Vs, respectively. The limits of detection (LDs) for K+, Na+ and Li+ ranged from 18 to 23µmolL−1. HD injection through an electronic micropipette allows to automatically dispense a bias-free amount of sample inside microchannels with acceptable repeatability. The proposed approach also exhibited instrumental simplicity, portability and minimal microfabrication requirements.

Is microrheometry affected by channel deformation?

Microrheometry is very important for exploring rheological behaviours of several systems when conventional techniques fail. Microrheometrical measurements are usually carried out in microfluidic devices made of Poly(dimethylsiloxane) (PDMS). Although PDMS is a very cheap material, it is also very easy to deform. In particular, a liquid flowing in a PDMS device, in some circumstances, can effectively deform the microchannel, thus altering the flow conditions. The measure of the fluid relaxation time might be performed through viscoelasticity induced particle migration in microfluidics devices. If the channel walls are deformed by the flow, the resulting measured value of the relaxation time could be not reliable. In this work, we study the effect of channel deformation on particle migration in square-shaped microchannel. Experiments are carried out in several PolyEthylene Oxyde solutions flowing in two devices made of PDMS and Poly(methylmethacrylate) (PMMA). The relevance of wall rigidity on particle migration is investigated, and the corresponding importance of wall rigidity on the determination of the relaxation time of the suspending liquid is examined.

Selective droplet sampling using a minimum number of horizontal pneumatic actuators in a high aspect ratio and highly flexible PDMS device

This paper presents a droplet sampling device driven by horizontal pneumatic actuators.

Rapid prototyping method for 3D PDMS microfluidic devices using a red femtosecond laser

A rapid prototyping technique is demonstrated which uses a red femtosecond laser to produce a metallic mould which is then directly used for the replica moulding of PDMS. The manufacturing process can be completed in less than 6 h making it a viable technique for testing new designs quickly. The technique is validated by creating a microfluidic device with channels of height and depth of 300 µm, with a ramp test structure where the height and width of the channels reduces to 100 µm to demonstrate the techniques 3D capabilities. The resulting PDMS device was easily removed from the metallic mould and closely replicated the shape aside the expected shrinkage during thermal curing. As the technique uses a single replica process, the surface roughness at the base of the channels corresponds to the un-ablated polished metal mould, resulting in a very low surface roughness of 0.361 nm. The ablated metallic mould surface corresponds to the top of the PDMS device, which is bonded to glass and does not affect the flow within the channels, reducing the need for optimisation of laser parameters. Finally, the device is validated by demonstrating laminar flow with the no-slip condition.

Fabrication of rigid microstructures with thiol-ene-based soft lithography for continuous-flow cell lysis.

In this work, we introduce a method for the soft-lithography-based fabrication of rigid microstructures and a new, simple bonding technique for use as a continuous-flow cell lysis device. While on-chip cell lysis techniques have been reported previously, these techniques generally require a long on-chip residence time, and thus cannot be performed in a rapid, continuous-flow manner. Microstructured microfluidic devices can perform mechanical lysis of cells, enabling continuous-flow lysis; however, rigid silicon-based devices require complex and expensive fabrication of each device, while polydimethylsiloxane (PMDS), the most common material used for soft lithography fabrication, is not rigid and expands under the pressures required, resulting in poor lysis performance. Here, we demonstrate the fabrication of microfluidic microstructures from off-stoichiometry thiol-ene (OSTE) polymer using soft-lithography replica molding combined with a post-assembly cure for easy bonding. With finite element simulations, we show that the rigid microstructures generate an energy dissipation rate of nearly 10(7), which is sufficient for continuous-flow cell lysis. Correspondingly, with the OSTE device we achieve lysis of highly deformable MDA-MB-231 breast cancer cells at a rate of 85%, while a comparable PDMS device leads to a lysis rate of only 40%.