Microfluidic Materials Research Articles

Collagen microsphere production on a chip

We have developed an integrated microfluidic material processing chip and demonstrated the rapid production of collagen microspheres encapsulating cells with high uniformity and cell viability. The chip integrated three material processing steps. Monodisperse microdroplets were generated at a microfluidic T junction between aqueous and mineral oil flows. The flow was heated immediately to 37 °C to initiate collagen fiber assembly within a gelation channel. Gelled microspheres were extracted from the mineral oil phase into cell culture media within an extraction chamber. Collagen gelation immediately after microdroplet generation significantly reduced coalescence among microdroplets that led to non-uniform microsphere production. The microfluidic extraction approach led to higher microsphere recovery and cell viability than when a conventional centrifugation extraction approach was employed. These results indicate that chip-based material processing is a promising approach for cell-ECM microenvironment generation for applications such as tissue engineering and stem cell delivery.

Inhibitory effect of common microfluidic materials on PCR outcome

In this study, we established a simple method for evaluating the PCR compatibility of various common materials employed when fabricating microfluidic chips, including silicon, several kinds of silicon oxide, glasses, plastics, wax, and adhesives. Two-temperature PCR was performed with these materials to determine their PCR-inhibitory effect. In most cases, adding bovine serum albumin effectively improved the reaction yield. We also studied the individual PCR components from the standpoint of adsorption. Most of the materials did not inhibit the DNA, although they noticeably interacted with the polymerase. We provide a simple method of performing PCR-compatibility testing of materials using inexpensive instrumentation that is common in molecular biology laboratories. Furthermore, our method is direct, being performed under actual PCR conditions with high temperature. Our results provide an overview of materials that are PCR-friendly for fabricating microfluidic devices. The PCR reaction, without any additives, performed best with pyrex glass, and it performed worst with PMMA or acrylic glue materials.

Single-Monomer Formulation of Polymerized Polyethylene Glycol Diacrylate as a Nonadsorptive Material for Microfluidics

Nonspecific adsorption in microfluidic systems can deplete target molecules in solution and prevent analytes, especially those at low concentrations, from reaching the detector. Polydimethylsiloxane (PDMS) is a widely used material for microfluidics, but it is prone to nonspecific adsorption, necessitating complex chemical modification processes to address this issue. An alternative material to PDMS that does not require subsequent chemical modification is presented here. Poly(ethylene glycol) diacrylate (PEGDA) mixed with photoinitiator forms on exposure to ultraviolet (UV) radiation a polymer with inherent resistance to nonspecific adsorption. Optimization of the polymerized PEGDA (poly-PEGDA) formula imbues this material with some of the same properties, including optical clarity, water stability, and low background fluorescence, that make PDMS so popular. Poly-PEGDA demonstrates less nonspecific adsorption than PDMS over a range of concentrations of flowing fluorescently tagged bovine serum albumin solutions, and poly-PEGDA has greater resistance to permeation by small hydrophobic molecules than PDMS. Poly-PEGDA also exhibits long-term (hour scale) resistance to nonspecific adsorption compared to PDMS when exposed to a low (1 μg/mL) concentration of a model adsorptive protein. Electrophoretic separations of amino acids and proteins resulted in symmetrical peaks and theoretical plate counts as high as 4 × 10(5)/m. Poly-PEGDA, which displays resistance to nonspecific adsorption, could have broad use in small volume analysis and biomedical research.

Whole-Teflon microfluidic chips.

Although microfluidics has shown exciting potential, its broad applications are significantly limited by drawbacks of the materials used to make them. In this work, we present a convenient strategy for fabricating whole-Teflon microfluidic chips with integrated valves that show outstanding inertness to various chemicals and extreme resistance against all solvents. Compared with other microfluidic materials [e.g., poly(dimethylsiloxane) (PDMS)] the whole-Teflon chip has a few more advantages, such as no absorption of small molecules, little adsorption of biomolecules onto channel walls, and no leaching of residue molecules from the material bulk into the solution in the channel. Various biological cells have been cultured in the whole-Teflon channel. Adherent cells can attach to the channel bottom, spread, and proliferate well in the channels (with similar proliferation rate to the cells in PDMS channels with the same dimensions). The moderately good gas permeability of the Teflon materials makes it suitable to culture cells inside the microchannels for a long time.

Hybrid Ceramic Polymers: New, Nonbiofouling, and Optically Transparent Materials for Microfluidics

A new, commercial hybrid ceramic polymer, Ormocomp, was introduced to the fabrication of microfluidic separation chips using two independent techniques, UV lithography and UV embossing. Both fabrication methods provided Ormocomp chips with stable cathodic electroosmotic flow which enabled examination of the Ormocomp biocompatibility by means of microchip capillary electrophoresis (MCE) and (intrinsic) fluorescence detection. The hydrophobic/hydrophilic properties of Ormocomp were examined by screening its interactions with bovine serum albumin and selected amino acids of varying hydrophobicity. The results show that the ceramic, organic-inorganic polymer structure natively resists biofouling on microchannel walls even so that the Ormocomp microchips can be used in intact protein analysis without prior surface modification. With theoretical separation plates approaching 10(4) m(-1) for intact proteins and 10(6) m(-1) for amino acids and peptides, our results suggest that Ormocomp microchips hold record-breaking performance as microfluidic separation platforms. In addition, Ormocomp was shown to be suitable for optical fluorescence detection even at near-UV range (ex 355 nm) with detection limits at a nanomolar level ( approximately 200 nM) for selected inherently fluorescent pharmaceuticals.

Cyclic olefin polymers: emerging materials for lab-on-a-chip applications

Cyclic olefin polymers (COPs) are increasingly popular as substrate material for microfluidics. This is due to their promising properties, such as high chemical resistance, low water absorption, good optical transparency in the near UV range and ease of fabrication. COPs are commercially available from a range of manufacturers under various brand names (Apel, Arton, Topas, Zeonex and Zeonor). Some of these (Apel and Topas) are made from more than one kind of monomer and therefore also known as cyclic olefin copolymers (COCs). In order to structure these materials, a wide array of fabrication methods is available. Laser ablation and micromilling are direct structuring methods suitable for fast prototyping, whilst injection moulding, hot embossing and nanoimprint lithography are replication methods more appropriate for low-cost production. Using these fabrication methods, a multitude of chemical analysis techniques have already been implemented. These include microchip electrophoresis (MCE), chromatography, solid phase extraction (SPE), isoelectric focusing (IEF) and mass spectrometry (MS). Still much additional work is needed to characterise and utilise the full potential of COP materials. This is especially true within optofluidics, where COPs are still rarely used, despite their excellent optical properties. This review presents a detailed description of the properties of COPs, the available fabrication methods and several selected applications described in the literature.

One-step fabrication of microfluidic chips with in-plane, adhesive-free interconnections

A simple method for creating interconnections to a common microfluidic device material, poly(methyl methacrylate) (PMMA), is presented. A press-fit interconnection is created between oversized, deformable tubing and complementary, undersized semi-circular ports fabricated into PMMA bonding surfaces by direct micromilling. Upon UV-assisted bonding the tubing is trapped in the ports of the PMMA chip and forms an integrated, in-plane and adhesive-free interconnection. The interconnections support the average pressure of 6.1 bar and can be made with small dead volumes. A comparison is made to a similar interconnection approach which uses tubing to act as a gasket between a needle and port on the microfluidic chip.

Synthesis and application of photo curable perfluoropolyethers as new material for microfluidics

We will describe the use of an alternative polymer which is suitable for use in microfluidic applications which require the use of harsh chemicals or aggressive solvents. Currently microfluidics systems are mainly manufactured in polydimethylsiloxane (PDMS) due to its ease of use, and the ability to produce monolithic microfluidic valves directly into the same material as the microfluidic chip. However, PDMS has limitations with respect to its chemical stability as it swells in contact with a lot of solvents and tends to become brittle when exposed to substances such as tetradecane. Polymers synthesized from perfluoropolyethers (PFPE) have significant advantages due to their outstanding chemical stability. Furthermore it is possible to photo cure these polymers allowing the convenient structuring of these polymers via lithography. We will demonstrate first experimental result in producing microfluidic components and their application with these polymers.

A study of the autofluorescence of parylene materials for μTAS applications

Parylene-C has been widely used as a biocompatible material for microfluidics and micro total analysis system (microTAS) applications in recent decades. However, its autofluorescence can be a major obstacle for parylene-C based devices used in applications requiring sensitive fluorescence detection. In this paper, Parylene-C was compared with other commonly used polymer and plastic materials in microTAS devices for their autofluorescence. We also report here an in-depth study of the behaviors and mechanisms of the autofluorescence of parylene-C, as well as several other commercialized members in the parylene family, including parylene-D, parylene-N and parylene-HT, using epifluorescence microscopy, fluorimeter and infrared spectroscopy. Strong autofluorescence was induced in parylene-C during short-wavelength excitation (i.e. UV excitation). Variation of autofluorescence intensity of parylene-C film was found to be related to both dehydrogenation and photo-oxidation. Moreover, the influence of microfabrication process on parylene-C autofluorescence was also evaluated. Parylene-HT, which exhibits low initial autofluorescence, decreasing autofluorescence behavior under UV excitation and higher UV stability, can be a promising alternative for microTAS applications with fluorescence detection.

Functional endothelialized microvascular networks with circular cross-sections in a tissue culture substrate

Functional endothelialized networks constitute a critical building block for vascularized replacement tissues, organ assist devices, and laboratory tools for in vitro discovery and evaluation of new therapeutic compounds. Progress towards realization of these functional artificial vasculatures has been gated by limitations associated with the mechanical and surface chemical properties of commonly used microfluidic substrate materials and by the geometry of the microchannels produced using conventional fabrication techniques. Here we report on a method for constructing microvascular networks from polystyrene substrates commonly used for tissue culture, built with circular cross-sections and smooth transitions at bifurcations. Silicon master molds are constructed using an electroplating process that results in semi-circular channel cross-sections with smoothly varying radii. These master molds are used to emboss polystyrene sheets which are then joined to form closed bifurcated channel networks with circular cross-sections. The mechanical and surface chemical properties of these polystyrene microvascular network structures enable culture of endothelial cells along the inner lumen. Endothelial cell viability was assessed, documenting nearly confluent monolayers within 3D microfabricated channel networks with rounded cross-sections.

Enabling Organosilicon Chemistries on Inert Polymer Surfaces with a Vapor-Deposited Silica Layer

Given the large surface area-to-volume ratios commonly encountered in microfluidics applications, the ability to engineer the chemical properties of surfaces encountered in these applications is critically important. However, as various polymers are rapidly replacing glass and silicon as the chosen materials for microfluidics devices, the ability to easily modify the surface chemistry has been diminished by the relatively inert nature of some commonly employed polymer surfaces, such as poly(methyl methacrylate) (PMMA), polystyrene, and polydimethylsiloxane (PDMS). This paper describes the low-temperature, vapor-phase deposition of robust silica layers to PMMA, polystyrene, and PDMS surfaces, which enables the functionalization of these surfaces by standard organosilane chemistries. Attenuated total reflection infrared spectroscopy, contact angle goniometry, ellipsometry, and atomic force microscopy are used to characterize the silica layers that form on these surfaces. Aqueous immersion experiments indicate that the silica layer has excellent stability in aqueous environments, which is a prerequisite for microfluidics applications, but for PMMA surfaces, low adhesion of the silica layer to the underlying substrate is problematic. For PDMS substrates, the presence of the silica layer helps to slow the process of hydrophobic recovery, which is an additional advantage.

Design of MEMS devices with optical apertures for the detection of transparent biological cells

This paper provides a novel technique to detect transparent biological living cells trapped in a microfluidic MEMS device by optical diffraction. The device essentially consists of an optical aperture or an aperture array patterned in metal layer and a microfluidic chamber positioned above the center of the aperture. When the cells in the chamber are illuminated through the aperture, the far-field diffraction pattern can be recorded by a CCD camera or a photodetector array. This diffraction pattern uniquely corresponds to the sizes, positions, and intrinsic optical properties of the aperture, cells, and the microfluidic chamber materials, so any unknown but relevant parameter is able to be extrapolated when all other parameters are fixed or identified. This paper describes in detail the designs of various microfluidic chambers and apertures for this application, and the development of a complete set of software for the analysis of the cells' optical properties. Compared with other currently available methods for the detection of transparent living cells, this method has the advantages of simple device structure, easy to manipulate, able to simultaneously detect several cells of different species, as well as providing accurate and sensitive results. Besides the detection of living cells, this technique can also be used to detect or characterize other transparent or low optical absorption particles, such as polymer spheres or insoluble droplets, inside an aqueous solution.

SU‐8 as a structural material for labs‐on‐chips and microelectromechanical systems

Since its introduction in the nineties, the negative resist SU-8 has been increasingly used in micro- and nanotechnologies. SU-8 has made the fabrication of high-aspect ratio structures accessible to labs with no high-end facilities such as X-ray lithography systems or deep reactive ion etching systems. These low-cost techniques have been applied not only in the fabrication of metallic parts or molds, but also in numerous other micromachining processes. Its ease of use has made SU-8 to be used in many applications, even when high-aspect ratios are not required. Beyond these pattern transfer applications, SU-8 has been used directly as a structural material for microelectromechanical systems and microfluidics due to its properties such as its excellent chemical resistance or the low Young modulus. In contrast to conventional resists, which are used temporally, SU-8 has been used as a permanent building material to fabricate microcomponents such as cantilevers, membranes, and microchannels. SU-8-based techniques have led to new low-temperature processes suitable for the fabrication of a wide range of objects, from the single component to the complete lab-on-chip. First, this article aims to review the different techniques and provides guidelines to the use of SU-8 as a structural material. Second, practical examples from our respective labs are presented.

Microfluidic protein patterning on silicon nitride using solvent-extracted poly(dimethylsiloxane) channels

Biomolecular patterning is essential for the creation of sensing motifs that rely on receptor-ligand binding for selectivity. Microfluidic devices have the potential to aid in the development of simple, robust methods for biomolecular patterning and therefore contribute to the generation of protein, DNA, and cell microarrays. In microfluidic patterning, the choice of both substrate and microfluidic channel material is essential for control of both the receptor binding for maximal signal generation as well as non-specific adsorption that acts as chemical noise. In this study, polystyrene, glass, silicon nitride, and poly(dimethylsiloxane) (PDMS) were evaluated as substrates for protein patterning using two types of PDMS microchannels for patterning, native PDMS and solvent-extracted PDMS (E-PDMS). E-PDMS microfluidic channels resulted in better patterning characteristics than native PDMS channels as determined by a higher fluorescence intensity of immobilized protein on all substrate types tested. Microfluidic patterning was then applied to perform two- and four-layer immunoassays.

Food Analysis on Microfluidic Devices Using Ultrasensitive Carbon Nanotubes Detectors

Microfluidic devices using carbon nanotube (CNT) materials (single-walled and two multiwalled (MWCNT)) for the analysis of selected analyte groups of significance in foods such as dietary antioxidants, water-soluble vitamins, vanilla flavors, and isoflavones involved in representative food samples have been explored for the first time. Ultrafast separations resulted in well-defined and resolved peaks with enhanced voltammetric current in comparison with those obtained from unmodified screen-printed electrodes, turning MWCNT into an ideal material for electrochemical sensing in food analysis. Resolution was improved by a factor of 2, and sensitivity was dramatically enhanced with amplification factors toward calibration slopes from 4- to 16-fold. In both qualitative and quantitative domains, this impressive performance of CNTs integrated on microfluidics allowed solving specific challenges in food environments such as the direct detection of analytes in complex natural samples and unambiguous analytes in the control of fraud, which was not possible on nonmodified surfaces, avoiding the integration of complex preconcentration steps on these microdevices. The use of these unique materials in microfluidics for food analysis has opened new expectations in "lab-on-a-chip" domains.

Real-time analysis of PCR inhibition on microfluidic materials

In order to clarify the suitability of different materials for development of microchip format PCR chambers, a real-time polymerase chain reaction (PCR) analysis was performed. The analysis was demonstrated on a modified 264 bp (base pair) PCR template fragment originating from the human GAPD gene, using SYBR Green® chemistry on different types of oxidized silicon, native silicon and other surfaces of interest. The PCR reactions with the test surfaces were studied to determine the inhibitory potential of the surfaces by comparing them to the reference reactions without the test surfaces. The analysis included the final level of fluorescence, the detection threshold delay, the amplification efficiency and the melting point analysis. Minor inhibition effects were observed for native silicon and SF6 etched surface without oxidation, while fluorocarbon-coated silicon showed a strong inhibition of PCR. Chemical, physical, as-deposited and thermal silicon dioxide surfaces on silicon resulted only in a delayed detection threshold, showing some level of inhibition during the early part of PCR. The chemically oxidized and the plasma oxidized silicon and silica surfaces were also analyzed with X-ray photoelectron spectrometer equipment showing the adsorption of DNA and the used polymerase. The effect of bovine serum albumine and poly-vinyl pyrrolidone in alleviating the inhibition was observed to be insignificant. Also the effect of starting amount of the template DNA was minimal, suggesting that the adsorption of DNA was not important for the observed inhibition.

Characteristics and impact of Taq enzyme adsorption on surfaces in microfluidic devices

We investigated the adsorption of Taq enzyme, using sessile droplets, on different microfluidic materials. In propagating adsorption materials, the contact angle (CA) of a sessile Taq droplet continually recedes and collapses due to adsorption. Contrastingly, in contained adsorption materials it exhibits an initial reduced CA due to an instantaneous adsorption, however remains time-invariant. Spectrophotometer analysis on SU8, a propagating adsorption material, reveals a gradual loss of Taq from the droplet onto the surface during droplet collapsing, as opposed to a rapid saturated adsorption in Teflon, a contained adsorption material. AFM micrographs of the adsorbed surfaces suggest a network-like structure in SU8 and distinctly different pillar-like structures in Teflon. With this understanding, we have successfully applied a SU8-Teflon coating to impart a time-invariant contact angle with minimal loss of Taq in surface microfluidic devices.

Photosensitive poly(dimethylsiloxane) materials for microfluidic applications

Poly(dimethylsiloxane) (PDMS) is used as a thermally crosslinked material in microfluidics and Bio-MEMS. Recently photo-patternable materials show increasing interest, as the demand for easy alignment arises for multilayered structures. We present a photopatterning process for PDMS in microfluidics, for two main uses: (a) as a thin (approximately 10 μm) structural layer, and (b) as a very thin (approximately 1 μm) hard mask for oxygen plasma etching of microfluidic polymeric substrates down to several tens of microns. We study the deep-UV and I-line photocrosslinking properties of siloxane copolymers containing vinyl-methyl-siloxane groups as polymerizable units. These materials are sensitive to DUV and can be sensitized to 300–400 nm using free radical initiators. We prove that even thermally curable PDMS (Sylgard 184, base) can become photosensitive in DUV, although its practical use is limited to very thin films, due to its small molecular weight.

Generation of Hydrophilic Poly(dimethylsiloxane) for High-Performance Microchip Electrophoresis

Poly(dimethylsiloxane) (PDMS) has become one of the most widely used materials for microchip capillary electrophoresis and microfluidics. The popularity of this material is the result of its low cost, simple fabrication, and rugged elastomeric properties. The hydrophobic nature of PDMS, however, limits its applicability for microchip CE, microfluidic patterning, and other nonelectrophoresis applications. The surface of PDMS can be made hydrophilic using a simple air plasma treatment; however, this property is quickly lost through hydrophobic recovery caused by diffusion of unreacted oligomer to the surface. Here, a simple approach for the generation of hydrophilic PDMS with long-term stability in air is presented. PDMS is rendered hydrophilic through a simple two-step extraction/oxidation process. First, PDMS is extracted in a series of solvents designed to remove unreacted oligomers from the bulk phase. Second, the oligomer-free PDMS is oxidized in a simple air plasma, generating a stable layer of hydrophilic SiO2. The conversion of surface-bound siloxane to SiO2 was followed with X-ray photoelectron spectroscopy. SiO2 on extracted-oxidized PDMS was stable for 7 days in air as compared to less than 3 h for native PDMS. Furthermore, the contact angle for modified PDMS was reduced to <40 degrees and remained low throughout the experiments. As a result of the decreased contact angle, capillary channels self-wet through capillary action, making the microchannels much easier to fill. Finally, the modification significantly improved the performance of the devices for microchip electrophoresis. The electroosmotic flow increased from 4.1 x 10(-4) to 6.8 x 10(-4) cm(2)/V.s for native compared to oxidized PDMS. Separation efficiencies for electrochemical detection also increased from 50 000 to 400 000 N/m for a 1.1-nL injection volume. The result of this modification is a significant improvement in the performance of PDMS for microchip electrophoresis and microfluidic applications.

Cross-Talk Problem on a Fluorescence Multi-Channel Microfluidic Chip System

Development of a compact fluorescence-based detection system for use in a micro-analytical system, such as a point-of-care diagnostic system, often requires a multi-channel microfluidic chip system. Since the materials used for microfluidic chips usually are transparent in the visible region and have a refractive indices higher than that of air or the surrounding environment, the fluorescence emission and scattered excitation light can propagate through the chip. We observed that such propagation can cause cross-talk between adjacent channels, and may become the major source of noise in the system and/or photo bleach the fluorescent samples in the adjacent channels, particularly for the small distances between the channels found in microfluidic chips, usually in order of several micro m. We monitored this cross-talk using fluorescein as a fluorescent sample and Mylar sheeting as a microfluidic chip material. We then discuss how this cross-talk can be avoided using a simple, inexpensive and effective method.