3D Microfluidic Networks Research Articles

Programmable microfluidic flow for automatic multistep digital assay in a single-sheet 3-dimensional paper-based microfluidic device

A single-sheet 3-dimensional paper-based microfluidic analytical device (3D-μPAD) has a user-friendly compact format that eliminates any potential for user alignment error compared to the stacking and folding format. However, achieving sophisticated analysis is still a challenge because forming a complex 3D microfluidic network sufficient for performing analysis in a single sheet of paper is difficult. Here, we present the first report of automatic multistep digital analysis in a single-sheet 3D-μPAD created by controlling the penetration of different wax inks into paper. Simple printing of different wax inks with different spreading properties produces different dimension microchannels in paper. This controllable change in the dimensions of the microchannels results in different flow rates of fluid in each microchannel. Based on the spatial resolution of the microchannels, our strategy enables programming of microfluidic flow through manipulation of the fluidic time delay. This approach creates 3D-μPADs capable of sequential fluid delivery and multistep biochemical reactions with single loading of a sample solution. The methodology can be further expanded to combination with digital assays to measure the concentration of a target analyte by simply counting the number of colored bars at a fixed time. It does not require any external instruments to analyze the measurement of the target analyte concentration. The significance of this work lies in the promising potential for automatic multistep digital assays in a low-cost and easy-to-use paper-based format that can be integrated with a roll-to-roll process for commercial-scale manufacturing.

In Situ Visualization and SERS Monitoring of the Interaction between Tumor and Endothelial Cells Using 3D Microfluidic Networks.

A multifunctional microfluidic platform was demonstrated to monitor the interaction between tumor cells and endothelial cells by integrating a three-dimensional (3D) cell culture unit with a protein detection unit. In such a chip, breast cancer cells MCF7 were seeded into the collagen to form a 3D tumor environment while human umbilical vein endothelial cells (HUVECs) are seeded in the channel next to the collagen matrix. Thus, an in situ growth of angiogenic sprouting can be visualized through fluorescence in the 3D collagen matrix after a coculture of MCF7 and HUVEC after 4 days, which cannot be observed in the 2D culture environment. On the other hand, gold@silver core-shell nanorods were used as surface-enhanced Raman scattering (SERS) immunoprobes for the detection of the secretion of cytokine (vascular endothelial growth factor, VEGF). The limit of detection of the VEGF is 100 pg/mL. Further, as LiCl and bevacizumab can act as a promoter and an inhibitor of VEGF, the dynamic change of the concentration of VEGF under the stimulation of them was monitored by SERS signals. Thus, this integrated SERS microfluidic platform creates opportunity for the fundamental research of interaction between tumors and endothelial cells.

Polymeric fully inertial lab-on-a-chip with enhanced-throughput sorting capabilities

In biology and medicine, the application of microfluidics filtration technologies to the separation of rare particles requires processing large amounts of liquid in a short time to achieve an effective separation yield. In this direction, the parallelization of the sorting process is desirable, but not so easy to implement in a lab on a chip (LoC) device, especially if it is fully inertial. In this work, we report on femtosecond laser microfabrication (FLM) of a poly(methyl methacrylate) (PMMA) inertial microfluidic sorter, separating particles based on their size and providing an enhanced-throughput capability. The LoC device consists of a microchannel with expansion chambers provided with siphoning outlets, for a continuous sorting process. Different from soft lithography, which is the most used technique for LoC prototyping, FLM allows developing 3D microfluidic networks connecting both sides of the chip. Exploiting this capability, we are able to parallelize the circuit while keeping a single output for the sorted particles and one for the remaining sample, thus increasing the number of processed particles per unit time without compromising the simplicity of the chip connections. We investigated several device layouts (at different flow rates) to define a configuration that maximizes the selectivity and the throughput.

Particle focusing by 3D inertial microfluidics

Three-dimensional (3D) particle focusing in microfluidics is a fundamental capability with a wide range of applications, such as on-chip flow cytometry, where high-throughput analysis at the single-cell level is performed. Currently, 3D focusing is achieved mainly in devices with complex layouts, additional sheath fluids, and complex pumping systems. In this work, we present a compact microfluidic device capable of 3D particle focusing at high flow rates and with a small footprint, without the requirement of external fields or lateral sheath flows, but using only a single-inlet, single-outlet microfluidic sequence of straight channels and tightly curving vertical loops. This device exploits inertial fluidic effects that occur in a laminar regime at sufficiently high flow rates, manipulating the particle positions by the combination of inertial lift forces and Dean drag forces. The device is fabricated by femtosecond laser irradiation followed by chemical etching, which is a simple two-step process enabling the creation of 3D microfluidic networks in fused silica glass substrates. The use of tightly curving three-dimensional microfluidic loops produces strong Dean drag forces along the whole loop but also induces an asymmetric Dean flow decay in the subsequent straight channel, thus producing rapid cross-sectional mixing flows that assist with 3D particle focusing. The use of out-of-plane loops favors a compact parallelization of multiple focusing channels, allowing one to process large amounts of samples. In addition, the low fluidic resistance of the channel network is compatible with vacuum driven flows. The resulting device is quite interesting for high-throughput on-chip flow cytometry.

A truly Lego®-like modular microfluidics platform

Ideally, a modular microfluidics platform should be simple to assemble and support 3D configurations for increased versatility. The modular building blocks should also be mass producible like electrical components. These are fundamental features of world-renowned Legos® and why Legos® inspire many existing modular microfluidics platforms. In this paper, a truly Lego®-like microfluidics platform is introduced, and its basic feasibility is demonstrated. Here, PDMS building blocks resembling 2 × 2 Lego® bricks are cast from 3D-printed master molds. The blocks are pegged and stacked on a traditional Lego® plate to create simple, 3D microfluidic networks, such as a single basket weave. Characteristics of the platform, including reversible sealing and automatic alignment of channels, are also analyzed and discussed in detail.

Bonding of SU-8 films onto KMPR structures for microfluidic, air-suspended photonic and optofluidic applications

We present a method to bond unstructured and structured SU-8 films down to sub-micron thicknesses onto microchannels fabricated in KMPR using a flexible polydimethylsiloxane (PDMS) stamp. By exploiting differently casted PDMS stamps, 3D microfluidic channel networks, air-suspended photonic devices and optofluidic structures have been fabricated. First, microchannels of KMPR are patterned by photolithography and an SU-8 film is spin coated onto a prepared PDMS stamp. The stamp is then placed on top of the KMPR microchannels and the SU-8 layer is cross-linked by applying sufficient heat and pressure. After peeling off the PDMS stamp, the SU-8 layer remains bonded on the KMPR. In our experiments, we demonstrate the bonding of approximately 0.5 μm thick structured SU-8 films onto KMPR microchannels of about 500 μm width and 25 μm height. Bond strength tests demonstrated that such thin layers can withstand pressures up to 1100 hPa. The laminated SU-8 layers can enable various functionalities, e.g. sealing of microfluidic channels, realization of air-suspended photonic structures or optofluidic devices. Most importantly, the combination of fluid handling in the microchannels and air-suspended photonic structures realized in the laminated SU-8 layer enables research towards a large range of applications, such as optofluidics, biosensors, chemical and biomedical analysis, environmental investigations, and renewable energy.

Leak-tight vertical membrane microvalves.

Pneumatic microvalves are fundamental control components in a large range of microfluidic applications. Their key performance parameters are small size, i.e. occupying a minimum of microfluidic real estate, low flow resistance in the open state, and leak-tight closing at limited control pressures. In this work we present the successful design, realization and evaluation of the first leak-tight, vertical membrane, pneumatic microvalves. The realization of the vertical membrane microvalves is enabled by a novel dual-sided molding method for microstructuring monolithic 3D microfluidic networks in PDMS in a single step, eliminating the need for layer-to-layer alignment during bonding. We demonstrate minimum lateral device features down to 20-30 μm in size, and vertical via density of ∼30 000 per cm(2), which provides significant gains in chip real estate compared to previously reported PDMS manufacturing methods. In contrast to horizontal membrane microvalves, there are no manufacturing restrictions on the cross-sectional geometry of the flow channel of the vertical membrane microvalves. This allows tuning the design towards lower closing pressure or lower open state flow resistance compared to those of horizontal membrane microvalves.

Construction of programmable interconnected 3D microfluidic networks

Microfluidic systems represent a key-enabling platform for novel diagnostic tools for use at the point-of-care in clinical contexts as well as for evolving single cell diagnostics. The design of 3D microfluidic systems is an active field of development, but construction of true interconnected 3D microfluidic networks is still a challenge, in particular when the goal is rapid prototyping, accurate design and flexibility. We report a novel approach for the construction of programmable 3D microfluidic systems consisting of modular 3D template casting of interconnected threads to allow user-programmable flow paths and examine its structural characteristics and its modular function. To overcome problems with thread template casting reported in the literature, low-surface-energy polymer threads were used, that allow solvent-free production. Connected circular channels with excellent roundness and low diameter variability were created. Variable channel termination allowed programming a flow path on-the-fly, thus rendering the resulting 3D microfluidic systems highly customizable even after production. Thus, construction of programmable/reprogrammable fully 3D microfluidic systems by template casting of a network of interconnecting threads is feasible, leads to high-quality and highly reproducible, complex 3D geometries.

Direct, one-step molding of 3D-printed structures for convenient fabrication of truly 3D PDMS microfluidic chips

In this work, we developed a convenient, one-step soft-lithographic-based molding technique for molding truly 3D microfluidic channels in polydimethylsiloxane (PDMS) by overcoming two grand challenges. We optimized the post-treatment conditions for 3D-printed resin structures to facilitate the use of them as masters for PDMS replica molding. What is more important, we demonstrated a novel method for single-step molding from 3D-printed microstructures to generate truly 3D microfluidic networks easily. With this technique, we fabricated some key, functional 3D microfluidic structures and components including a basket-weaving network, a 3D chaotic advective mixer and microfluidic peristaltic valves. Furthermore, an interesting “injection-on-demand” microfluidic device was also demonstrated. Our technique offers a simple, fast route to the fabrication of 3D microfluidic chips in a short time without clean-room facilities.

Manufacturing of Three-dimensionally Microstructured Nanocomposites through Microfluidic Infiltration

Microstructured composite beams reinforced with complex three-dimensionally (3D) patterned nanocomposite microfilaments are fabricated via nanocomposite infiltration of 3D interconnected microfluidic networks. The manufacturing of the reinforced beams begins with the fabrication of microfluidic networks, which involves layer-by-layer deposition of fugitive ink filaments using a dispensing robot, filling the empty space between filaments using a low viscosity resin, curing the resin and finally removing the ink. Self-supported 3D structures with other geometries and many layers (e.g. a few hundreds layers) could be built using this method. The resulting tubular microfluidic networks are then infiltrated with thermosetting nanocomposite suspensions containing nanofillers (e.g. single-walled carbon nanotubes), and subsequently cured. The infiltration is done by applying a pressure gradient between two ends of the empty network (either by applying a vacuum or vacuum-assisted microinjection). Prior to the infiltration, the nanocomposite suspensions are prepared by dispersing nanofillers into polymer matrices using ultrasonication and three-roll mixing methods. The nanocomposites (i.e. materials infiltrated) are then solidified under UV exposure/heat cure, resulting in a 3D-reinforced composite structure. The technique presented here enables the design of functional nanocomposite macroscopic products for microengineering applications such as actuators and sensors.

Manufacturing of Three-dimensionally Microstructured Nanocomposites through Microfluidic Infiltration

Microstructured composite beams reinforced with complex three-dimensionally (3D) patterned nanocomposite microfilaments are fabricated via nanocomposite infiltration of 3D interconnected microfluidic networks. The manufacturing of the reinforced beams begins with the fabrication of microfluidic networks, which involves layer-by-layer deposition of fugitive ink filaments using a dispensing robot, filling the empty space between filaments using a low viscosity resin, curing the resin and finally removing the ink. Self-supported 3D structures with other geometries and many layers (e.g. a few hundreds layers) could be built using this method. The resulting tubular microfluidic networks are then infiltrated with thermosetting nanocomposite suspensions containing nanofillers (e.g. single-walled carbon nanotubes), and subsequently cured. The infiltration is done by applying a pressure gradient between two ends of the empty network (either by applying a vacuum or vacuum-assisted microinjection). Prior to the infiltration, the nanocomposite suspensions are prepared by dispersing nanofillers into polymer matrices using ultrasonication and three-roll mixing methods. The nanocomposites (i.e. materials infiltrated) are then solidified under UV exposure/heat cure, resulting in a 3D-reinforced composite structure. The technique presented here enables the design of functional nanocomposite macroscopic products for microengineering applications such as actuators and sensors.

Flock‐Based Microfluidics: Flock‐Based Microfluidics (Adv. Mater. 19/2013)

Electrostatic flocking is a technique for depositing vertically-aligned microfibers onto an adhesive-coated substrate using a high electrostatic field. Emmanuel Delamarche and co-workers use this technique on page 2672 to pattern layers of flock fibers resulting in 2D and 3D microfluidic networks, which fill spontaneously due to capillary action. With this approach, multifunctional microfluidic platforms for diagnostic applications can be produced at extremely low cost.

Layer-by-layer micromolding of natural biopolymer scaffolds with intrinsic microfluidic networks

A three-dimensional (3D) microfluidic network plays an important role in engineering thick organs. However, most of the existing methods are limited to mechanically robust synthetic biomaterials and only planar or simple microfluidic networks have been incorporated into soft natural biopolymers. Here we presented an automatic layer-by-layer micromolding strategy to reproducibly fabricate 3D microfluidic porous scaffolds directly from the aqueous solution of soft natural biopolymers. Process parameters such as the liquid volume for each layer and contact displacement were investigated to produce a structurally stable 3D microfluidic scaffold. Microscopic characterization demonstrated that the microfluidic channels were interconnected in 3D and successfully functioned as a convective pathway to transport a polymer solution. Endothelial cells grew relatively well in the porous microfluidic channels. It is envisioned that this method could provide an alternative way to reproducibly build complex 3D microfluidic networks into extracellular matrix-like scaffolds for the fabrication of soft vascularized organs.

Construction of microfluidic biochips with enhanced functionalities using 3D femtosecond laser direct writing

The extreme nonlinear interaction betweenfemtosecond laser pulses and large-band-gapmaterials has enabled three-dimensional (3D)microfabrication inside transparent materials. In thepast decade, this technique has been used forcreating a variety of functional components in glassmaterials, including microoptics, microfluidics,microelectronics, micromechanics, etc. Using thesebuilding blocks, femtosecond laser microfabricationalso allows for construction of highly integratedmicrodevices. Here, we provide an overview of ourlatest progress made along this direction, includingfocal spot engineering and nanofluidic fabrication.In particular, we show that 3D micro-/nano-fluidiccomponents with arbitrary geometries can bedirectly formed inside glass. This opens uppromising prospects for a broad spectrum ofapplications based on compact and complex 3Dmicrofluidic networks. Our work shows that thistechnique holds promise for fabricating 3D hybridmicro-systems, such as Lab-on-a-chip devices andMicro Total Analysis Systems in the future.

Bottom-up fabrication of 3D cell-laden microfluidic constructs

Microfluidic scaffold or hydrogel has attracted extensive interests in generating thick tissues or organs. It is still challenging to produce complex three-dimensional (3D) microfluidic network from natural biomaterials like silk fibroin and chitosan. Here we present a bottom-up approach to fabricate 3D microfluidic constructs by rolling monolayer cell-loaded scaffolds. The results demonstrated that the microfluidic channels were maintained relatively well and the cells were uniformly distributed in 3D. The 3D cell-laden microfluidic constructs were dynamically cultured in a house-made bioreactor system to enhance mass transport and the cells inside exhibited long-term viability. We envision that this simple yet versatile method can be potentially used to engineer 3D vascularized tissues.

A 3D Interconnected Microchannel Network Formed in Gelatin by Sacrificial Shellac Microfibers

3D microfluidic networks are fabricated in a gelatin hydrogel using sacrificial melt-spun microfibers made from a material with pH-dependent solubility. The fibers, after being embedded within the gel, can be removed by changing the gel pH to induce dissolution. This process is performed in an entirely aqueous environment, avoiding extreme temperatures, low pressures, and toxic organic solvents.

Chemistry with spatial control using particles and streams().

Spatial control of chemical reactions, with micro- and nanometer scale resolution, has important consequences for one pot synthesis, engineering complex reactions, developmental biology, cellular biochemistry and emergent behavior. We review synthetic methods to engineer this spatial control using chemical diffusion from spherical particles, shells and polyhedra. We discuss systems that enable both isotropic and anisotropic chemical release from isolated and arrayed particles to create inhomogeneous and spatially patterned chemical fields. In addition to such finite chemical sources, we also discuss spatial control enabled with laminar flow in 2D and 3D microfluidic networks. Throughout the paper, we highlight applications of spatially controlled chemistry in chemical kinetics, reaction-diffusion systems, chemotaxis and morphogenesis.

System-level simulation of liquid filling in microfluidic chips

Liquid filling in microfluidic channels is a complex process that depends on a variety of geometric, operating, and material parameters such as microchannel geometry, flow velocity∕pressure, liquid surface tension, and contact angle of channel surface. Accurate analysis of the filling process can provide key insights into the filling time, air bubble trapping, and dead zone formation, and help evaluate trade-offs among the various design parameters and lead to optimal chip design. However, efficient modeling of liquid filling in complex microfluidic networks continues to be a significant challenge. High-fidelity computational methods, such as the volume of fluid method, are prohibitively expensive from a computational standpoint. Analytical models, on the other hand, are primarily applicable to idealized geometries and, hence, are unable to accurately capture chip level behavior of complex microfluidic systems. This paper presents a parametrized dynamic model for the system-level analysis of liquid filling in three-dimensional (3D) microfluidic networks. In our approach, a complex microfluidic network is deconstructed into a set of commonly used components, such as reservoirs, microchannels, and junctions. The components are then assembled according to their spatial layout and operating rationale to achieve a rapid system-level model. A dynamic model based on the transient momentum equation is developed to track the liquid front in the microchannels. The principle of mass conservation at the junction is used to link the fluidic parameters in the microchannels emanating from the junction. Assembly of these component models yields a set of differential and algebraic equations, which upon integration provides temporal information of the liquid filling process, particularly liquid front propagation (i.e., the arrival time). The models are used to simulate the transient liquid filling process in a variety of microfluidic constructs and in a multiplexer, representing a complex microfluidic network. The accuracy (relative error less than 7%) and orders-of-magnitude speedup (30 000X-4 000 000X) of our system-level models are verified by comparison against 3D high-fidelity numerical studies. Our findings clearly establish the utility of our models and simulation methodology for fast, reliable analysis of liquid filling to guide the design optimization of complex microfluidic networks.

Interfacing Three-Dimensional Curved Structures and Cellular Adhesion

Curvature is a fundamental geometric design principle found in an array of biological systems, such as vasculature. Therefore, in studying cellular processes such as adhesion, proliferation and migration, it is important to consider the effects of curved 3D micro-topography as compared to flat 2D substrates, which are far more common. Creating these 3D curved systems requires novel approaches as well. Microfluidic devices would seem to be a good approach for this as they have often been utilized as a platform for studying cell adhesion and migration in vitro. However, the fabrication of curved, non-rectangular channels has been a major challenge to the field of microfluidics due to conventional fabrication methods. To overcome these limitations, we have developed a novel and robust approach using mechanical micromachining in combination with a two-step reverse polymer molding process to fabricate microfluidic channels with circular cross-sectional geometries. Here, we utilize these 3D microfluidic networks to study the effects of curvature on cell adhesion mechanics. Both fibroblast (NIH-3T3) and endothelial (HUVEC) cell lines were cultured within circular cross-section microfluidic channels and on reserve molded cylindrical curved polymers. Cell morphology on these curved versus flat substrates was then characterized via confocal and scanning electron microscopy. Furthermore, the formation of stable focal adhesions and cytoskeletal organization was analyzed by immunofluorescent confocal microscopy. We believe that this approach for fabricating bioinspired microfluidic systems provides a powerful platform for interfacing cellular interactions with curved 3D structures, which could be useful in a variety of fields from vascular biology and immune cell transmigration to cell mechanotransduction and tissue engineering.

Capillary-driven automatic packaging

Packaging continues to be one of the most challenging steps in micro-nanofabrication, as many emerging techniques (e.g., soft lithography) are incompatible with the standard high-precision alignment and bonding equipment. In this paper, we present a simple-to-operate, easy-to-adapt packaging strategy, referred to as Capillary-driven Automatic Packaging (CAP), to achieve automatic packaging process, including the desired features of spontaneous alignment and bonding, wide applicability to various materials, potential scalability, and direct incorporation in the layout. Specifically, self-alignment and self-engagement of the CAP process induced by the interfacial capillary interactions between a liquid capillary bridge and the top and bottom substrates have been experimentally characterized and theoretically analyzed with scalable implications. High-precision alignment (of less than 10 µm) and outstanding bonding performance (up to 300 kPa) has been reliably obtained. In addition, a 3D microfluidic network, aligned and bonded by the CAP technique, has been devised to demonstrate the applicability of this facile yet robust packaging technique for emerging microfluidic and bioengineering applications.