共查询到20条相似文献，搜索用时 15 毫秒
1.
Here, we present a simple chemical modification of poly(dimethylsiloxane) (PDMS) by curing a mixture of 2 wt% undecylenic acid (UDA) in PDMS prepolymer on a gold-coated glass slide. This gold slide had been previously pretreated with a self-assembled hydrophilic monolayer of 3-mercaptopropionic acid (MPA). During curing of the UDA∕PDMS prepolymer, the hydrophilic UDA carboxyl moieties diffuses toward the hydrophilic MPA carboxyl moieties on the gold surface. This diffusion of the UDA within the PDMS prepolymer to the surface is a direct result of surface energy minimization. Once completely cured, the PDMS is peeled off the gold substrate, thereby exposing the interfacial carboxyl groups. These groups are then available for subsequent attachment of 5(')-amino terminated DNA oligonucleotides via amide linkages. Our results show that the covalently tethered oligonucleotides can successfully capture fluorescein-labeled complementary oligonucleotides via hybridization, which are visualized using fluorescence microscopy. 相似文献
2.
Systematic characterization of degas-driven flow for poly(dimethylsiloxane) microfluidic devices 总被引：1，自引：0，他引：1
Degas-driven flow is a novel phenomenon used to propel fluids in poly(dimethylsiloxane) (PDMS)-based microfluidic devices without requiring any external power. This method takes advantage of the inherently high porosity and air solubility of PDMS by removing air molecules from the bulk PDMS before initiating the flow. The dynamics of degas-driven flow are dependent on the channel and device geometries and are highly sensitive to temporal parameters. These dependencies have not been fully characterized, hindering broad use of degas-driven flow as a microfluidic pumping mechanism. Here, we characterize, for the first time, the effect of various parameters on the dynamics of degas-driven flow, including channel geometry, PDMS thickness, PDMS exposure area, vacuum degassing time, and idle time at atmospheric pressure before loading. We investigate the effect of these parameters on flow velocity as well as channel fill time for the degas-driven flow process. Using our devices, we achieved reproducible flow with a standard deviation of less than 8% for flow velocity, as well as maximum flow rates of up to 3 nL∕s and mean flow rates of approximately 1-1.5 nL∕s. Parameters such as channel surface area and PDMS chip exposure area were found to have negligible impact on degas-driven flow dynamics, whereas channel cross-sectional area, degas time, PDMS thickness, and idle time were found to have a larger impact. In addition, we develop a physical model that can predict mean flow velocities within 6% of experimental values and can be used as a tool for future design of PDMS-based microfluidic devices that utilize degas-driven flow. 相似文献
3.
Cell culture and harvest are the most upstream operation for a completely integrated cell assay chip. In our previous work, thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm) was successfully grafted onto polydimethylsiloxane (PDMS) surface via benzophenone-initiated photopolymerization. In the present work, the PNIPAAm-grafted-PDMS (PNIPAAm-g-PDMS) surface was explored for thermomodulated cell culture and noninvasive harvest in microfluidic channels. Using COS 7 fibroblast from African green monkey kidney as the model cells, the thermomodulated adhering and detaching behaviors of the cells on the PNIPAAm-g-PDMS surfaces were optimized with respect to PNIPAAm-grafting yields and gelatin modification. The viability of the cells cultured on and harvested from the PNIPAAm-g-PDMS surface with the thermomodulated noninvasive protocol was estimated against the traditional cell culture∕harvest method involving trypsin digestion. The configuration of the microchannel on the PNIPAAm-g-PDMS chip was evaluated for static cell culture. Using a pipette-shaped PNIPAAm-g-PDMS microchannel, long-term cell culture could be achieved at 37 °C with periodic change of the culture medium every 12 h. After moving the microchip from the incubator set at 37 °C to the room temperature, the proliferated cells could be spontaneously detached from the PNIPAAm-g-PDMS surface of the upstream chamber and transferred by a gentle fluid flow to the downstream chamber, wherein the transferred cells could be subcultured. The thermomodulated cell culture, harvest, and passage operations on the PNIPAAm-g-PDMS microfluidic channels were demonstrated. 相似文献
4.
A new method of surface modification is described for enabling the in situ formation of homogenous porous polymer monoliths (PPMs) within poly(dimethylsiloxane) (PDMS) microfluidic channels that uses 365 nm UV illumination for polymerization. Porous polymer monolith formation in PDMS can be challenging because PDMS readily absorbs the monomers and solvents, changing the final monolith morphology, and because PDMS absorbs oxygen, which inhibits free-radical polymerization. The new approach is based on sequentially absorbing a non-hydrogen-abstracting photoinitiator and the monomers methyl methacrylate and ethylene diacrylate within the walls of the microchannel, and then polymerizing the surface treatment polymer within the PDMS, entangled with it but not covalently bound. Four different monolith compositions were tested, all of which yielded monoliths that were securely anchored and could withstand pressures exceeding the bonding strength of PDMS (40 psi) without dislodging. One was a recipe that was optimized to give a larger average pore size, required for low back pressure. This monolith was used to concentrate and subsequently mechanical lyse B lymphocytes. 相似文献
5.
Definable surface chemistry is essential for many applications of microfluidic polymer systems. However, small cross-section channels with a high surface to volume ratio enhance passive adsorption of molecules that depletes active molecules in solution and contaminates the channel surface. Here, we present a one-step photochemical process to coat the inner surfaces of closed microfluidic channels with a nanometer thick layer of poly(ethylene glycol) (PEG), well known to strongly reduce non-specific adsorption, using only commercially available reagents in an aqueous environment. The coating consists of PEG diacrylate (PEGDA) covalently grafted to polymer surfaces via UV light activation of the water soluble photoinitiator benzoyl benzylamine, a benzophenone derivative. The PEGDA coating was shown to efficiently limit the adsorption of antibodies and other proteins to <5% of the adsorbed amount on uncoated polymer surfaces. The coating could also efficiently suppress the adhesion of mammalian cells as demonstrated using the HT-29 cancer cell line. In a subsequent equivalent process step, protein in aqueous solution could be anchored onto the PEGDA coating in spatially defined patterns with a resolution of <15 μm using an inverted microscope as a projection lithography system. Surface patterns of the cell binding protein fibronectin were photochemically defined inside a closed microfluidic device that was initially homogeneously coated by PEGDA. The resulting fibronectin patterns were shown to greatly improve cell adhesion compared to unexposed areas. This method opens for easy surface modification of closed microfluidic systems through combining a low protein binding PEG-based coating with spatially defined protein patterns of interest. 相似文献
6.
A simple method for micromanipulation of liquids and∕or small groups of cells is presented in this study. Microfabricated sieving structures composed of PDMS (polydimethylsiloxane) were used to segregate aqueous solutions. This microfluidic valving scheme was an application of Cassie-Baxter wetting and was termed "virtual walls" as a nonsolid barrier exists at an air∕water interface. The manipulation of the virtual-air-wall valve was accomplished by controlling the strength of surface-tension and hydrostatic-pressure forces. Virtual walls with a range of feature sizes were designed and characterized by monitoring air and water displacement in response to hydrostatic pressure. Thresholds for the virtual-air-wall valves to be turned on or off were quantified. The walls could also be formed or dissipated by the focused microbeam of a pulsed laser. As an illustration of the virtual wall utility, a series of microfluidic applications were demonstrated. First, the capability of virtual walls to temporarily segregate liquids was integrated into a device utilized to establish a chemical gradient. In a second application, the arraying of nonadherent cells within individual aqueous cavities created by the virtual walls was demonstrated. Individual cells were also released from the cavities on demand using a focused microbeam. The virtual walls were simple and easy-to-fabricate without the requirement for surface treatment or precision alignment, and should find usage in bioanalytical applications. 相似文献
7.
Surface modification is a critical issue in various applications of polydimethylsiloxane (PDMS)-based microfluidic devices. Here, we describe a novel method through which PDMS-based microchannels were successfully modified with fragmented poly(l-lactic acid) (PLLA) nanosheets through a simple patchwork technique that exploited the high level of adhesiveness of PLLA nanosheets. Compared with other surface modification methods, our method required neither complicated chemical modifications nor the use of organic solvents that tend to cause PDMS swelling. The experimental results indicated that the modified PDMS exhibited excellent capacity for preventing the adhesion and activation of platelets. This simple yet efficient method can be used to fabricate the special PDMS microfluidic devices for biological, medical, and even hematological purposes. 相似文献
8.
Seamless integration of biological components with electrochemical sensors is critical in the development of microdevices for cell analysis. The present paper describes the integration miniature Au electrodes next to immune cells (macrophages) in order to detect cell-secreted hydrogen peroxide (H(2)O(2)). Photopatterning of poly(ethylene glycol) (PEG) hydrogels was used to both immobilize horseradish peroxidase molecules onto electrodes and to define regions for cell attachment in the vicinity of sensing electrodes. Electrodes micropatterned in such a manner were enclosed inside poly(dimethylsiloxane) fluid conduits and incubated with macrophages. The cells attached onto the exposed glass regions in the vicinity of the electrodes and nowhere else on the non-fouling PEG hydrogel surface. A microfluidic device was converted into an electrochemical cell by placing flow-through Ag∕AgCl reference and Pt wire counter electrodes at the outlet and inlet, respectively. This microdevice with integrated H(2)O(2)-sensing electrodes had sensitivity of 27 μA∕cm(2) mM with a limit of detection of 2 μM. Importantly, this microdevice allowed controllable seeding of macrophages next to electrodes, activation of these cells and on-chip monitoring of H(2)O(2) release in real time. In the future, this biosensor platform may be utilized for monitoring of macrophage responses to pathogens or for the study of inflammatory signaling in micropatterned cell cultures. 相似文献
9.
In this paper, one-step air plasma treatment is successfully used for poly(dimethylsiloxane)(PDMS)-plastic chip bonding. The technique is green, cheap, and requires no other reagent other than air. Hydrocarbon plastics: polystyrene (PS), cyclic olefin copolymer (COC), and polypropylene (PP) have all been successfully bonded to PDMS irreversibly. The corresponding compressed air resistances are measured to be around 500 kPa for PDMS-PS, PDMS-COC, and PDMS-PP hybrid chips. The bondings are also of good quality even after storage under different temperatures and subject to solutions from acid to base. 相似文献
10.
Vascular function, homeostasis, and pathological development are regulated by the endothelial cells that line blood vessels. Endothelial function is influenced by the integrated effects of multiple factors, including hemodynamic conditions, soluble and insoluble biochemical signals, and interactions with other cell types. Here, we present a membrane microfluidic device that recapitulates key components of the vascular microenvironment, including hemodynamic shear stress, circulating cytokines, extracellular matrix proteins, and multiple interacting cells. The utility of the device was demonstrated by measuring monocyte adhesion to and transmigration through a porcine aortic endothelial cell monolayer. Endothelial cells grown in the membrane microchannels and subjected to 20 dynes∕cm(2) shear stress remained viable, attached, and confluent for several days. Consistent with the data from macroscale systems, 25 ng∕ml tumor necrosis factor (TNF)-α significantly increased RAW264.7 monocyte adhesion. Preconditioning endothelial cells for 24 h under static or 20 dynes∕cm(2) shear stress conditions did not influence TNF-α-induced monocyte attachment. In contrast, simultaneous application of TNF-α and 20 dynes∕cm(2) shear stress caused increased monocyte adhesion compared with endothelial cells treated with TNF-α under static conditions. THP-1 monocytic cells migrated across an activated endothelium, with increased diapedesis in response to monocyte chemoattractant protein (MCP)-1 in the lower channel of the device. This microfluidic platform can be used to study complex cell-matrix and cell-cell interactions in environments that mimic those in native and tissue engineered blood vessels, and offers the potential for parallelization and increased throughput over conventional macroscale systems. 相似文献
11.
Precisely controlling the spatial distribution of biomolecules on biomaterial surface is important for directing cellular activities in the controlled cell microenvironment. This paper describes a polydimethylsiloxane (PDMS) gradient-generating microfluidic device to immobilize the gradient of cellular adhesive Arg-Gly-Asp (RGD) peptide on poly (ethylene glycol) (PEG) hydrogel. Hydrogels are formed by exposing the mixture of PEG diacrylate (PEGDA), acryloyl-PEG-RGD, and photo-initiator with ultraviolet light. The microfluidic chip was simulated by a fluid dynamic model for the biomolecule diffusion process and gradient generation. PEG hydrogel covalently immobilized with RGD peptide gradient was fabricated in this microfluidic device by photo-polymerization. Bone marrow derived rat mesenchymal stem cells (MSCs) were then cultured on the surface of RGD gradient PEG hydrogel. Cell adhesion of rat MSCs on PEG hydrogel with various RGD gradients were then qualitatively and quantitatively analyzed by immunostaining method. MSCs cultured on PEG hydrogel surface with RGD gradient showed a grated fashion for cell adhesion and spreading that was proportional to RGD concentration. It was also found that 0.107–0.143 mM was the critical RGD concentration range for MSCs maximum adhesion on PEG hydrogel. 相似文献
12.
Fang Ye Jin Jiang Honglong Chang Li Xie Jinjun Deng Zhibo Ma Weizheng Yuan 《Biomicrofluidics》2015,9(4)
Cell studies at the single-cell level are becoming more and more critical for understanding the complex biological processes. Here, we present an optimization study investigating the positioning of single cells using micromolding in capillaries technology coupled with the cytophobic biomaterial poly (2-hydroxyethyl methacrylate) (poly (HEMA)). As a cytophobic biomaterial, poly (HEMA) was used to inhibit cells, whereas the glass was used as the substrate to provide a cell adhesive background. The poly (HEMA) chemical barrier was obtained using micromolding in capillaries, and the microchannel networks used for capillarity were easily achieved by reversibly bonding the polydimethylsiloxane mold and the glass. Finally, discrete cell adhesion regions were presented on the glass surface. This method is facile and low cost, and the reagents are commercially available. We validated the cytophobic abilities of the poly (HEMA), optimized the channel parameters for higher quality and more stable poly (HEMA) patterns by investigating the effects of changing the aspect ratio and the width of the microchannel on the poly (HEMA) grid pattern, and improved the single-cell occupancy by optimizing the dimensions of the cell adhesion regions. 相似文献
13.
We have investigated the bonding stability of various silane treatments for the integration of
track-etched membranes with poly(dimethylsiloxane) (PDMS) microfluidic devices. We compare various
treatments using trialkoxysilanes or dipodal silanes to determine the effect of the organofunctional
group, cross-link density, reaction solvent, and catalyst on the bond stability. We find that
devices made using existing silane methods delaminated after one day when immersed in cell culture
medium at 37 °C. In contrast, the dipodal silane, bis[3-(trimethoxysilyl)propyl]amine, is shown to
yield stable and functional integration of membranes with PDMS that is suitable for long-term cell
culture. To demonstrate application of the technique, we fabricated an open-surface device in which
cells cultured on a track-etched membrane can be stimulated at their basal side via embedded
microfluidic channels. C2C12 mouse myoblasts were differentiated into myotubes over the course of
two weeks on these devices to demonstrate biocompatibility. Finally, devices were imaged during the
basal-side delivery of a fluorescent stain to validate the membrane operation and long-term
stability of the bonding technique. 相似文献
14.
Extracellular matrix (ECM) proteins are required for cell culture. In this paper, we report the use of O(2) plasma bonding to fabricate a perfusion culture microchamber array chip with identical-size ECM spots in the isolated microchambers. The chip was fabricated by assembly of two poly(dimethylsiloxane) (PDMS) layers, a microfluidic network layer, and an ECM array layer, which were aligned and then bonded by O(2) plasma oxidation with protection of the ECM microarray with a physical mask made from PDMS. We successfully cultivated Chinese hamster ovary K1 cells in the microchambers with fibronectin. In the fibronectin microchambers, the cells adhered and extended after 12 h of static culture and then grew over the course of 1 d of perfusion culture. 相似文献
15.
Hiroaki Takehara Akira Nagaoka Jun Noguchi Takanori Akagi Takamasa Sakai Ung-il Chung Haruo Kasai Takanori Ichiki 《Biomicrofluidics》2013,7(5)
Hydrogels have several excellent characteristics suitable for biomedical use such as softness, biological inertness and solute permeability. Hence, integrating hydrogels into microfluidic devices is a promising approach for providing additional functions such as biocompatibility and porosity, to microfluidic devices. However, the poor mechanical strength of hydrogels has severely limited device design and fabrication. A tetra-poly(ethylene glycol) (tetra-PEG) hydrogel synthesized recently has high mechanical strength and is expected to overcome such a limitation. In this research, we have comprehensively studied the implementation of tetra-PEG gel into microfluidic device technology. First, the fabrication of tetra-PEG gel/PDMS hybrid microchannels was established by developing a simple and robust bonding technique. Second, some fundamental features of tetra-PEG gel/PDMS hybrid microchannels, particularly fluid flow and mass transfer, were studied. Finally, to demonstrate the unique application of tetra-PEG-gel-integrated microfluidic devices, the generation of patterned chemical modulation with the maximum concentration gradient: 10% per 20 μm in a hydrogel was performed. The techniques developed in this study are expected to provide fundamental and beneficial methods of developing various microfluidic devices for life science and biomedical applications. 相似文献
16.
Cell-microscale pattern surface interactions are crucial to understand many fundamental biological questions and develop regenerative medicine and tissue engineering approaches. In this work, we demonstrated a simple method to pattern PDMS surface by sacrificing poly vinyl pyrrolidone (PVP) electrospinning nanofibers and investigated the growth profile of cells on the modified patterned surfaces using stroma cells. The stromal cells were observed to exhibit good viability on this modified surface and the patterned surface with alignment nanofibers could promote cell migration. Furthermore, the modified PDMS surface was integrated with microfluidic channels to create the microscale spatial factor and was used to explore the cell migration and orientation under this microsystem. Both spatial factor and patterned surfaces were found to contribute to the complex cell orientation under the combined dual effects. This established method is simple, fast, and easy for use, demonstrating the potential of this microsystem for applications in addressing biological questions in complex environment. 相似文献
17.
Katja Uhlig Bj?rn Boysen Andreas Lankenau Magnus Jaeger Erik Wischerhoff Jean-Fran?ois Lutz André Laschewsky Claus Duschl 《Biomicrofluidics》2012,6(2)
Thermoresponsive polymer surface coatings are a promising tool for cell culture applications. They allow for a mild way of cell detachment that preserves the activity of membrane proteins—a prerequisite for reliable cell analysis. To enlarge the application range of these coatings to cells with different adhesion properties, we synthesized various novel poly(ethylene glycol)-based thermoresponsive polymers and describe how (i) their chemical structure and (ii) their surface density affect their efficiency. In order to quantify the influence of both factors, the time for cell spreading and rounding efficiency were observed. As a result, efficiency of cell rounding, which is closely correlated to cell detachment, is less affected by both factors than the time needed for cell spreading. This time can effectively be adjusted by the molecular architecture which includes the length of the polymer backbone and the side chains. Based on this work, recommendations are given for future optimization of functionality of thermoresponsive polymer coatings for cell culture applications. 相似文献
18.
This paper reviews our work on the application of ultrafast pulsed laser micro∕nanoprocessing for the three-dimensional (3D) biomimetic modification of materials surfaces. It is shown that the artificial surfaces obtained by femtosecond-laser processing of Si in reactive gas atmosphere exhibit roughness at both micro- and nanoscales that mimics the hierarchical morphology of natural surfaces. Along with the spatial control of the topology, defining surface chemistry provides materials exhibiting notable wetting characteristics which are potentially useful for open microfluidic applications. Depending on the functional coating deposited on the laser patterned 3D structures, we can achieve artificial surfaces that are (a) of extremely low surface energy, thus water-repellent and self-cleaned, and (b) responsive, i.e., showing the ability to change their surface energy in response to different external stimuli such as light, electric field, and pH. Moreover, the behavior of different kinds of cells cultured on laser engineered substrates of various wettabilities was investigated. Experiments showed that it is possible to preferentially tune cell adhesion and growth through choosing proper combinations of surface topography and chemistry. It is concluded that the laser textured 3D micro∕nano-Si surfaces with controllability of roughness ratio and surface chemistry can advantageously serve as a novel means to elucidate the 3D cell-scaffold interactions for tissue engineering applications. 相似文献
19.
Walter Schrott Zdeněk Slouka Petr ?ervenka Ji?í Ston Marek Nebyla Michal P?ibyl Dalimil ?nita 《Biomicrofluidics》2009,3(4)
Electrokinetic properties and morphology of PDMS microfluidic chips intended for bioassays are studied. The chips are fabricated by a casting method followed by polymerization bonding. Microchannels are coated with 1% solution of bovine serum albumin (BSA) in Tris buffer. Albumin passively adsorbs on the PDMS surface. Electrokinetic characteristics (electro-osmotic velocity, electro-osmotic mobility, and zeta potential) of the coated PDMS channels are experimentally determined as functions of the electric field strength and the characteristic electrolyte concentration. Atomic force microscopy (AFM) analysis of the surface reveals a “peak and ridge” structure of the protein layer and an imperfect substrate coating. On the basis of the AFM observation, several topologies of the BSA-PDMS surface are proposed. A nonslip mathematical model of the electro-osmotic flow is then numerically analyzed. It is found that the electrokinetic characteristics computed for a channel with the homogeneous distribution of a fixed electric charge do not fit the experimental data. Heterogeneous distribution of the fixed electric charge and the surface roughness is thus taken into account. When a flat PDMS surface with electric charge heterogeneities is considered, the numerical results are in very good agreement with our experimental data. An optimization analysis finally allowed the determination of the surface concentration of the electric charge and the degree of the PDMS surface coating. The obtained findings can be important for correct prediction and possibly for robust control of behavior of electrically driven PDMS microfluidic chips. The proposed method of the electro-osmotic flow analysis at surfaces with a heterogeneous distribution of the surface electric charge can also be exploited in the interpretation of experimental studies dealing with protein-solid phase interactions or substrate coatings. 相似文献
20.
The development of widely applicable point-of-care sensing and diagnostic devices can benefit from simple and inexpensive fabrication techniques that expedite the design, testing, and implementation of lab-on-a-chip devices. In particular, electrodes integrated within microfluidic devices enable the use of electrochemical techniques for the label-free detection of relevant analytes. This work presents a novel, simple, and cost-effective bench-top approach for the integration of high surface area three-dimensional structured electrodes fabricated on polystyrene (PS) within poly(dimethylsiloxane) (PDMS)-based microfluidics. Optimization of PS-PDMS bonding results in integrated devices that perform well under pressure and fluidic flow stress. Furthermore, the fabrication and bonding processes are shown to have no effect on sensing electrode performance. Finally, the on-chip sensing capabilities of a three-electrode electrochemical cell are demonstrated with a model redox compound, where the high surface area structured electrodes exhibit ultra-high sensitivity. We propose that the developed approach can significantly expedite and reduce the cost of fabrication of sensing devices where arrays of functionalized electrodes can be used for point-of-care analysis and diagnostics. 相似文献