PDMS-based Microfluidic Devices Research Articles

Advancing Tissue Culture with Light-Driven 3D-Printed Microfluidic Devices.

Three-dimensional (3D) printing presents a compelling alternative for fabricating microfluidic devices, circumventing certain limitations associated with traditional soft lithography methods. Microfluidics play a crucial role in the biomedical sciences, particularly in the creation of tissue spheroids and pharmaceutical research. Among the various 3D printing techniques, light-driven methods such as stereolithography (SLA), digital light processing (DLP), and photopolymer inkjet printing have gained prominence in microfluidics due to their rapid prototyping capabilities, high-resolution printing, and low processing temperatures. This review offers a comprehensive overview of light-driven 3D printing techniques used in the fabrication of advanced microfluidic devices. It explores biomedical applications for 3D-printed microfluidics and provides insights into their potential impact and functionality within the biomedical field. We further summarize three light-driven 3D printing strategies for producing biomedical microfluidic systems: direct construction of microfluidic devices for cell culture, PDMS-based microfluidic devices for tissue engineering, and a modular SLA-printed microfluidic chip to co-culture and monitor cells.

Two-Photon Polymerization 3D-Printing of Micro-scale Neuronal Cell Culture Devices.

Neuronal cultures have been a reference experimental model for several decades. However, 3D cell arrangement, spatial constraints on neurite outgrowth, and realistic synaptic connectivity are missing. The latter limits the study of structure and function in the context of compartmentalization and diminishes the significance of cultures in neuroscience. Approximating ex vivo the structured anatomical arrangement of synaptic connectivity is not trivial, despite being key for the emergence of rhythms, synaptic plasticity, and ultimately, brain pathophysiology. Here, two-photon polymerization (2PP) is employed as a 3D printing technique, enabling the rapid fabrication of polymeric cell culture devices using polydimethyl-siloxane (PDMS) at the micrometer scale. Compared to conventional replica molding techniques based on microphotolitography, 2PP micro-scale printing enables rapid and affordable turnaround of prototypes. This protocol illustrates the design and fabrication of PDMS-based microfluidic devices aimed at culturing modular neuronal networks. As a proof-of-principle, a two-chamber device is presented to physically constrain connectivity. Specifically, an asymmetric axonal outgrowth during ex vivo development is favored and allowed to be directed from one chamber to the other. In order to probe the functional consequences of unidirectional synaptic interactions, commercial microelectrode arrays are chosen to monitor the bioelectrical activity of interconnected neuronal modules. Here, methods to 1) fabricate molds with micrometer precision and 2) perform in vitro multisite extracellular recordings in rat cortical neuronal cultures are illustrated. By decreasing costs and future widespread accessibility of 2PP 3D-printing, this method will become more and more relevant across research labs worldwide. Especially in neurotechnology and high-throughput neural data recording, the ease and rapidity of prototyping simplified in vitro models will improve experimental control and theoretical understanding of in vivo large-scale neural systems.

Superhydrophobic treatment of PDMS-based microfluidic devices using CO2 laser ablation

Superhydrophobic treatment of PDMS-based microfluidic devices using CO2 laser ablation

Direct electrification of silicon microfluidics for electric field applications

Microfluidic systems are widely used in fundamental research and industrial applications due to their unique behavior, enhanced control, and manipulation opportunities of liquids in constrained geometries. In micrometer-sized channels, electric fields are efficient mechanisms for manipulating liquids, leading to deflection, injection, poration or electrochemical modification of cells and droplets. While PDMS-based microfluidic devices are used due to their inexpensive fabrication, they are limited in terms of electrode integration. Using silicon as the channel material, microfabrication techniques can be used to create nearby electrodes. Despite the advantages that silicon provides, its opacity has prevented its usage in most important microfluidic applications that need optical access. To overcome this barrier, silicon-on-insulator technology in microfluidics is introduced to create optical viewports and channel-interfacing electrodes. More specifically, the microfluidic channel walls are directly electrified via selective, nanoscale etching to introduce insulation segments inside the silicon device layer, thereby achieving the most homogeneous electric field distributions and lowest operation voltages feasible across microfluidic channels. These ideal electrostatic conditions enable a drastic energy reduction, as effectively shown via picoinjection and fluorescence-activated droplet sorting applications at voltages below 6 and 15 V, respectively, facilitating low-voltage electric field applications in next-generation microfluidics.

Enhanced and stabilized mesenchymal stem cell growth inside plasma pre-treated and collagen-coated PDMS-based microfluidic chambers

Poly(dimethylsiloxane) (PDMS) – based microfluidic devices, let them be lab-on-a-chip or organ-on-a-chip platforms, have been extensively utilized in biomedical research, due to the versatility, rapid prototyping, optical transparency and ease of fabrication. Despite its numerous advantages, PDMS is inherently hydrophobic, which compromises cell adhesion and proliferation, given the direct correlation between surface properties and cell viability. Various strategies have been followed for the functionalization of PDMS surface, from which extracellular matrix (ECM) protein immobilization appears to be more compatible with cell-related applications. In this context, current studies propose multi-step, tedious and occasionally environmentally hazardous procedures of PDMS chemical functionalization that are additionally not easily employed in microfluidic systems. To overcome these limitations, this work suggests air plasma treatment as the single-step PDMS activation procedure for the immobilization of type I collagen, and is one of the few to actually be applied inside microfluidic devices, at the integral fabrication step of sealing, to study cell sustainability. Stable hydrophilization of the plasma-treated and collagen-coated PDMS surfaces was verified for a time period of 7 days in dry state, achieving the highest reported wettability. Moreover, mesenchymal stem cells (MSCs) were cultured in as-treated 3D microchambers for 5 days, when they reached complete confluency, one of the highest reported in literature. The results revealed the potential of plasma treatment followed shortly after by collagen layering to support stabilized and long-term cell culture in PDMS-based microfluidic devices. The herein proposed PDMS surface functionalization method can be used to create a stable PDMS-collagen composite layer in organ-on-a-chip platforms.

Recent Progress and Perspectives on Neural Chip Platforms Integrating PDMS-Based Microfluidic Devices and Microelectrode Arrays.

Recent years have witnessed a spurt of progress in the application of the encoding and decoding of neural activities to drug screening, diseases diagnosis, and brain-computer interactions. To overcome the constraints of the complexity of the brain and the ethical considerations of in vivo research, neural chip platforms integrating microfluidic devices and microelectrode arrays have been raised, which can not only customize growth paths for neurons in vitro but also monitor and modulate the specialized neural networks grown on chips. Therefore, this article reviews the developmental history of chip platforms integrating microfluidic devices and microelectrode arrays. First, we review the design and application of advanced microelectrode arrays and microfluidic devices. After, we introduce the fabrication process of neural chip platforms. Finally, we highlight the recent progress on this type of chip platform as a research tool in the field of brain science and neuroscience, focusing on neuropharmacology, neurological diseases, and simplified brain models. This is a detailed and comprehensive review of neural chip platforms. This work aims to fulfill the following three goals: (1) summarize the latest design patterns and fabrication schemes of such platforms, providing a reference for the development of other new platforms; (2) generalize several important applications of chip platforms in the field of neurology, which will attract the attention of scientists in the field; and (3) propose the developmental direction of neural chip platforms integrating microfluidic devices and microelectrode arrays.

Surface Optimization and Design Adaptation toward Spheroid Formation On-Chip.

Spheroids have become an essential tool in preclinical cancer research. The uniformity of spheroids is a critical parameter in drug test results. Spheroids form by self-assembly of cells. Hence, the control of homogeneity of spheroids in terms of size, shape, and density is challenging. We developed surface-optimized polydimethylsiloxane (PDMS) biochip platforms for uniform spheroid formation on-chip. These biochips were surface modified with 10% bovine serum albumin (BSA) to effectively suppress cell adhesion on the PDMS surface. These surface-optimized platforms facilitate cell self-aggregations to produce homogenous non-scaffold-based spheroids. We produced uniform spheroids on these biochips using six different established human cell lines and a co-culture model. Here, we observe that the concentration of the BSA is important in blocking cell adhesion to the PDMS surfaces. Biochips treated with 3% BSA demonstrated cell repellent properties similar to the bare PDMS surfaces. This work highlights the importance of surface modification on spheroid production on PDMS-based microfluidic devices.

Rapid Production of PDMS Microdevices for Electrodriven Separations and Microfluidics by 3D-Printed Scaffold Removal

In our work, we produced PDMS-based microfluidic devices by mechanical removal of 3D-printed scaffolds inserted in PDMS. Two setups leading to the fabrication of monolithic PDMS-based microdevices and bonded (or stamped) PDMS-based microdevices were designed. In the monolithic devices, the 3D-printed scaffolds were fully inserted in the PDMS and then carefully removed. The bonded devices were produced by forming imprints of the 3D-printed scaffolds in PDMS, followed by bonding the PDMS parts to glass slides. All these microfluidic devices were then successfully employed in three proof-of-concept applications: capture of magnetic microparticles, formation of droplets, and isotachophoresis separation of model organic dyes.

Characterization of Nanoparticle Adsorption on Polydimethylsiloxane-Based Microchannels

Nanoparticles (NPs) are used in various medicinal applications. Exosomes, bio-derived NPs, are promising biomarkers obtained through separation and concentration from body fluids. Polydimethylsiloxane (PDMS)-based microchannels are well-suited for precise handling of NPs, offering benefits such as high gas permeability and low cytotoxicity. However, the large specific surface area of NPs may result in nonspecific adsorption on the device substrate and thus cause sample loss. Therefore, an understanding of NP adsorption on microchannels is important for the operation of microfluidic devices used for NP handling. Herein, we characterized NP adsorption on PDMS-based substrates and microchannels by atomic force microscopy to correlate NP adsorptivity with the electrostatic interactions associated with NP and dispersion medium properties. When polystyrene NP dispersions were introduced into PDMS-based microchannels at a constant flow rate, the number of adsorbed NPs decreased with decreasing NP and microchannel zeta potentials (i.e., with increasing pH), which suggested that the electrostatic interaction between the microchannel and NPs enhanced their repulsion. When exosome dispersions were introduced into PDMS-based microchannels with different wettabilities at constant flow rates, exosome adsorption was dominated by electrostatic interactions. The findings obtained should facilitate the preconcentration, separation, and sensing of NPs by PDMS-based microfluidic devices.

Surface Modification of PDMS-Based Microfluidic Devices with Collagen Using Polydopamine as a Spacer to Enhance Primary Human Bronchial Epithelial Cell Adhesion.

Polydimethylsiloxane (PDMS) is a silicone-based synthetic material used in various biomedical applications due to its properties, including transparency, flexibility, permeability to gases, and ease of use. Though PDMS facilitates and assists the fabrication of complicated geometries at micro- and nano-scales, it does not optimally interact with cells for adherence and proliferation. Various strategies have been proposed to render PDMS to enhance cell attachment. The majority of these surface modification techniques have been offered for a static cell culture system. However, dynamic cell culture systems such as organ-on-a-chip devices are demanding platforms that recapitulate a living tissue microenvironment’s complexity. In organ-on-a-chip platforms, PDMS surfaces are usually coated by extracellular matrix (ECM) proteins, which occur as a result of a physical and weak bonding between PDMS and ECM proteins, and this binding can be degraded when it is exposed to shear stresses. This work reports static and dynamic coating methods to covalently bind collagen within a PDMS-based microfluidic device using polydopamine (PDA). These coating methods were evaluated using water contact angle measurement and atomic force microscopy (AFM) to optimize coating conditions. The biocompatibility of collagen-coated PDMS devices was assessed by culturing primary human bronchial epithelial cells (HBECs) in microfluidic devices. It was shown that both PDA coating methods could be used to bind collagen, thereby improving cell adhesion (approximately three times higher) without showing any discernible difference in cell attachment between these two methods. These results suggested that such a surface modification can help coat extracellular matrix protein onto PDMS-based microfluidic devices.

Conventional and emerging strategies for the fabrication and functionalization of PDMS-based microfluidic devices

Microfluidics is an emerging and multidisciplinary field that is of great interest to manufacturers in medicine, biotechnology, and chemistry, as it provides unique tools for the development of point-of-care diagnostics, organs-on-chip systems, and biosensors. Polymeric microfluidics, unlike glass and silicon, offer several advantages such as low-cost mass manufacturing and a wide range of beneficial material properties, which make them the material of choice for commercial applications and high-throughput systems. Among polymers used for the fabrication of microfluidic devices, polydimethylsiloxane (PDMS) still remains the most widely used material in academia due to its advantageous properties, such as excellent transparency and biocompatibility. However, commercialization of PDMS has been a challenge mostly due to the high cost of the current fabrication strategies. Moreover, specific surface modification and functionalization steps are required to tailor the surface chemistry of PDMS channels (e.g. biomolecule immobilization, surface hydrophobicity and antifouling properties) with respect to the desired application. While significant research has been reported in the field of PDMS microfluidics, functionalization of PDMS surfaces remains a critical step in the fabrication process that is difficult to navigate. This review first offers a thorough illustration of existing fabrication methods for PDMS-based microfluidic devices, providing several recent advancements in this field with the aim of reducing the cost and time for mass production of these devices. Next, various conventional and emerging approaches for engineering the surface chemistry of PDMS are discussed in detail. We provide a wide range of functionalization techniques rendering PDMS microchannels highly biocompatible for physical or covalent immobilization of various biological entities while preventing non-specific interactions.

Time Dependent Lyotropic Chromonic Textures in Microfluidic Confinements

Nematic and columnar phases of lyotropic chromonic liquid crystals (LCLCs) have been long studied for their fundamental and applied prospects in material science and medical diagnostics. LCLC phases represent different self-assembled states of disc-shaped molecules, held together by noncovalent interactions that lead to highly sensitive concentration and temperature dependent properties. Yet, microscale insights into confined LCLCs, specifically in the context of confinement geometry and surface properties, are lacking. Here, we report the emergence of time dependent textures in static disodium cromoglycate (DSCG) solutions, confined in PDMS-based microfluidic devices. We use a combination of soft lithography, surface characterization, and polarized optical imaging to generate and analyze the confinement-induced LCLC textures and demonstrate that over time, herringbone and spherulite textures emerge due to spontaneous nematic (N) to columnar M-phase transition, propagating from the LCLC-PDMS interface into the LCLC bulk. By varying the confinement geometry, anchoring conditions, and the initial DSCG concentration, we can systematically tune the temporal dynamics of the N- to M-phase transition and textural behavior of the confined LCLC. Overall, the time taken to change from nematic to the characteristic M-phase textures decreased as the confinement aspect ratio (width/depth) increased. For a given aspect ratio, the transition to the M-phase was generally faster in degenerate planar confinements, relative to the transition in homeotropic confinements. Since the static molecular states register the initial conditions for LC flows, the time dependent textures reported here suggest that the surface and confinement effects—even under static conditions—could be central in understanding the flow behavior of LCLCs and the associated transport properties of this versatile material.

Endothelialization of PDMS-based microfluidic devices under high shear stress conditions

Microfluidic systems made out of polydimethylsiloxane (PDMS) offer a platform to mimic vascular flow conditions in model systems at well-defined shear stresses. However, extracellular matrix (ECM) proteins that are physisorbed on the PDMS are not reliably attached under high shear stress conditions, which makes long-term experiments difficult. To overcome this limitation, we functionalized PDMS surfaces with 3-aminopropyltriethoxysilane (APTES) by using different surface activation methods to develop a stable linkage between the PDMS surface and collagen, which served as a model ECM protein. The stability of the protein coating inside the microfluidic devices was evaluated in perfusion experiments with phosphate-buffered saline (PBS) at 10−40 dynes/cm2 wall shear stress. To assess the stability of cell adhesion, endothelial cells were grown in a multi-shear device over a shear stress range of 20−150 dynes/cm2. Cells on the APTES-mediated collagen coating were stable over the entire shear stress range in PBS (pH 9) for 48 h. The results suggest that at high pH values, the electrostatic interaction between APTES-coated surfaces and collagen molecules offer a very promising tool to modify PDMS-based microfluidic devices for long-term endothelialization under high shear stress conditions.

Microwave irradiation-based rapid curing of PDMS for microfluidic applications

Microfluidic devices are routinely fabricated using replica moulding. The master relief structure is generally constructed on a silicon substrate and the stamps are made using polydimethylsiloxane (PDMS), cured via the slow process of hot-air convective heating. An alternative method that greatly reduces this cure time is presented here. It employs a low-cost, consumer-grade microwave oven and speeds up the curing process by permitting a much higher heat flux to be applied to the prepolymer. A specific pulsed irradiation protocol has been devised which achieves precisely controlled energy delivery for this purpose. This is necessary because silicon’s relatively much stronger microwave absorption causes uncontrollable temperature overshoots that could degrade PDMS if continuously irradiated. The protocol has been arrived at after due optimization of operational parameters, achieved via simulation of a finite element model. It has been implemented by modifying the control circuitry of the consumer grade oven. Cure times as low as two minutes have been demonstrated. Optical and mechanical characterization results show no adverse effect on the performance of PDMS cured using the proposed method. Thus, it can be used for the rapid fabrication of PDMS-based microfluidic devices and would be especially beneficial for research labs looking to speed up microfluidic device development iterations.

Universal method for fabricating PDMS microfluidic device using SU8, 3D printing and soft lithography

Microfluidic devices are constructed from polydimethylsiloxane (PDMS) due to their biocompatibility, fabrication ease, well-established protocols, and simplicity. PDMS-based microfluidic devices are constructed by (i) applying liquid PDMS to a negative mold (usually a silicon or 3D-printed mold) and (ii) curing the PDMS with heat exposure over a set time period. Unreacted resin monomers in 3D-printed molds prevent PDMS from fully curing, resulting in improper channel formation in PDMS and reducing the PDMS device’s efficacy. An in-house protocol that uses SU-8 as a “non-stick” coating on 3D-printed molds facilitates the successful casting of PDMS. Contact angle, surface profile, optical profile, and force testing prove that PDMS cast from SU-8-treated molds resembles pristine PDMS, unlike PDMS cast from untreated molds. Further, this method is generalized to commercial 3D prints using different 3D printing resins. To demonstrate this technique’s viability in microfluidic devices, a microfluidic tree using PDMS from treated 3D prints shows vibrant colors and clear lines. This is absent from an untreated PDMS.

PDMS-based microfluidic devices with shrinkable wax molds printed on biaxially orientated polystyrene film

This study proposed a novel method for the fabrication of shrinkable wax mold for PDMS-based microfluidic devices. In this approach, the designed wax patterns were printed on the surface of a thermally shrinkable polymer film (biaxially orientated polystyrene) with a commercial wax printer. Then a thermal treatment process was conducted, which simultaneously triggered wax reflow and shrinkage of the polymer film. After cooling, finer wax patterns with increased aspect ratio were achieved. Finally, wax patterns on polymer film can be used as the mold for the casting of PDMS-based microfluidic devices. Compared with previous approaches for the fabrication of molds for PDMS, the proposed fabrication method is more rapid, low-cost, non-toxic and without the requirement of cleanroom facilities or photolithography instruments. In addition, the use of thermally shrinkable polymer substrate helps to achieve an increased aspect ratio, as well as finer and higher wax patterns. And the wax patterns could be used as the mold for the fabrication of PDMS-based microfluidics.

Resolving protein mixtures using microfluidic diffusional sizing combined with synchrotron radiation circular dichroism.

Circular dichroism spectroscopy has become a powerful tool to characterise proteins and other biomolecules. For heterogeneous samples such as those present for interacting proteins, typically only average spectroscopic features can be resolved. Here we overcome this limitation by using free-flow microfluidic size separation in-line with synchrotron radiation circular dichroism to resolve the secondary structure of each component of a model protein mixture containing monomers and fibrils. To enable this objective, we have integrated far-UV compatible measurement chambers into PDMS-based microfluidic devices. Two architectures are proposed so as to accommodate for a wide range of concentrations. The approach, which can be used in combination with other bulk measurement techniques, paves the way to the study of complex mixtures such as the ones associated with protein misfolding and aggregation diseases including Alzheimer's and Parkinson's diseases.

PDMS-Based Microfluidic Devices for Cell Culture

Microfluidic technology has affirmed itself as a powerful tool in medical and biological research by offering the possibility of managing biological samples in tiny channels and chambers. Among the different applications, the use of microfluidics for cell cultures has attracted much interest from scientists worldwide. Traditional cell culture methods need high quantities of samples and reagents that are strongly reduced in miniaturized systems. In addition, the microenvironment is better controlled by scaling down. In this paper, we provide an overview of the aspects related to the design of a novel microfluidic culture chamber, the fabrication approach based on polydimethylsiloxane (PDMS) soft-lithography, and the most critical issues in shrinking the size of the system.

Viable cell culture in PDMS-based microfluidic devices.

Microfluidics has played a vital role in developing novel methods to investigate biological phenomena at the molecular and cellular level during the last two decades. Microscale engineering of cellular systems is nevertheless a nascent field marked inherently by frequent disruptive advancements in technology such as PDMS-based soft lithography. Viable culture and manipulation of cells in microfluidic devices requires knowledge across multiple disciplines including molecular and cellular biology, chemistry, physics, and engineering. There has been numerous excellent reviews in the past 15 years on applications of microfluidics for molecular and cellular biology including microfluidic cell culture (Berthier et al., 2012; El-Ali, Sorger, & Jensen, 2006; Halldorsson et al., 2015; Kim et al., 2007; Mehling & Tay, 2014; Sackmann et al., 2014; Whitesides, 2006; Young & Beebe, 2010), cell culture models (Gupta et al., 2016; Inamdar & Borenstein, 2011; Meyvantsson & Beebe, 2008), cell secretion (Schrell et al., 2016), chemotaxis (Kim & Wu, 2012; Wu et al., 2013), neuron culture (Millet & Gillette, 2012a, 2012b), drug screening (Dittrich & Manz, 2006; Eribol, Uguz, & Ulgen, 2016; Wu, Huang, & Lee, 2010), cell sorting (Autebert et al., 2012; Bhagat et al., 2010; Gossett et al., 2010; Wyatt Shields Iv, Reyes, & López, 2015), single cell studies (Lecault et al., 2012; Reece et al., 2016; Yin & Marshall, 2012), stem cell biology (Burdick & Vunjak-Novakovic, 2009; Wu et al., 2011; Zhang & Austin, 2012), cell differentiation (Zhang et al., 2017a), systems biology (Breslauer, Lee, & Lee, 2006), 3D cell culture (Huh et al., 2011; Li et al., 2012; van Duinen et al., 2015), spheroids and organoids (Lee et al., 2016; Montanez-Sauri, Beebe, & Sung, 2015; Morimoto & Takeuchi, 2013; Skardal et al., 2016; Young, 2013), organ-on-chip (Bhatia & Ingber, 2014; Esch, Bahinski, & Huh, 2015; Huh et al., 2011; van der Meer & van den Berg, 2012), and tissue engineering (Andersson & Van Den Berg, 2004; Choi et al., 2007; Hasan et al., 2014). In this chapter, we provide an overview of PDMS-based microdevices for microfluidic cell culture. We discuss the advantages and challenges of using PDMS-based soft lithography for microfluidic cell culture and highlight recent progress and future directions in this area.

PDMS-based microfluidic devices using commoditized PCBs as masters with no specialized equipment required

A simple, convenient and reliable approach used to prepare general polymer PDMS-based microfluidic devices with a minimal requirement for equipment.