Poly Dimethylsiloxane Microchip Research Articles

Highly efficient detection of insulinotropic action of glucagon via GLP-1 receptor in mice pancreatic beta-cell with a novel perfusion microchip

Abstract Glucagon exhibits insulinotropic ability by activating cAMP through glucagon or glucagon-like peptide-1 (GLP-1) receptors. To investigate the mechanism of endogenous and exogenous glucagon on insulin release, we studied the receptor selectivity on pancreatic islet beta-cells by switching the glucose concentration from 20 mmol/L to 0 mmol/L. To measure the exact temporal relationship between glucagon and insulin release, we developed a quick, small volume, multi-channel polydimethylsiloxane (PDMS) microchip. At 0 mmol/L glucose, we observed an insulinotropic effect in both INS-1 cells and islets. Meanwhile, we observed a 63 ± 6.27 s delay of endogenous glucagon-induced insulin release. After treatment with glucagon and GLP-1 receptor antagonists, we found that endogenous glucagon utilized the glucagon receptor, whereas exogenous glucagon primarily utilized the GLP-1 receptor to promote insulin secretion. The microchip can also be used to describe the “glucagonocentric” vision of diabetes pathophysiology. Taken together, the insulinotropic mechanism of different receptors should be taken into account in clinical treatments.

One-step preparation and application of mussel-inspired poly(norepinephrine)-coated polydimethylsiloxane microchip for separation of chiral compounds.

In this paper, using the self-polymerization of norepinephrine (NE) and its favorable film-forming property, a simple and green preparation approach was developed to modify a PDMS channel for enantioseparation of chiral compounds. After the PDMS microchip was filled with NE solution, poly(norepinephrine) (PNE) film was gradually formed and deposited on the inner wall of microchannel as permanent coating via the oxidation of NE by the oxygen dissolved in the solution. Due to possessing plentiful catechol and amine functional groups, the PNE-coated PDMS microchip exhibited much better wettability, more stable and suppressed EOF, and less nonspecific adsorption. The water contact angle and EOF of PNE-coated PDMS substrate were measured to be 13° and 1.68 × 10(-4) cm(2) V(-1) s(-1) , compared to those of 108° and 2.24 × 10(-4) cm(2) V(-1) s(-1) from the untreated one, respectively. Different kinds of chiral compounds, such as amino acid enantiomer, drug enantiomer, and peptide enantiomer were efficiently separated utilizing a separation length of 37 mm coupled with in-column amperometric detection on the PNE-coated PDMS microchips. This facile mussel-inspired PNE-based microchip system exhibited strong recognition ability, high-performance, admirable reproducibility, and stability, which may have potential use in the complex biological analysis.

Electrokinetic sample preconcentration and hydrodynamic sample injection for microchip electrophoresis using a pneumatic microvalve.

A microfluidic platform was developed to perform online electrokinetic sample preconcentration and rapid hydrodynamic sample injection for zone electrophoresis using a single microvalve. The polydimethylsiloxane microchip comprises a separation channel, a side channel for sample introduction, and a control channel which is used as a pneumatic microvalve aligned at the intersection of the two flow channels. The closed microvalve, created by multilayer soft lithography, serves as a nanochannel preconcentrator under an applied electric potential, enabling current to pass through while preventing bulk flow. Once analytes are concentrated, the valve is briefly opened and the stacked sample is pressure injected into the separation channel for electrophoretic separation. Fluorescently labeled peptides were enriched by a factor of ∼450 in 230 s. This method enables both rapid analyte concentration and controlled injection volume for high sensitivity, high-resolution CE.

A novel polydimethylsiloxane microfluidic viscometer fabricated using microwire-molding

We present a new economical microfluidic viscometer to measure the viscosity of biological fluids, using sample volumes of less than 200 μl. It is fabricated using a microwire-molding technique, making it easier and cheaper to produce than existing viscometers. The viscometer is based on laminar flow inside a polydimethylsiloxane microchip. The velocity of the sample flow inside the capillary was monitored with a camera, and the movement of the liquid column was determined by a Matlab video-processing program. The device was calibrated using deionized water, which is a Newtonian fluid, at 20 °C. The viscometer provides accurate measurements of viscosity for values as small as 0.69 mPa s. The viscosity of water at different temperatures was measured, showing more than 98% agreement with the values provided by the National Institute of Standards and Technology. Various samples including a series of glycerol solutions, phosphate-buffered saline, alcohol, and cell media were also tested, and the measured viscosities were compared with those from a traditional glass capillary viscometer. The results show good agreement between the two methods, with an average relative error of less than 1%. Furthermore, the viscosities of several cell suspensions were measured, showing a relative standard deviation of less than 1.5%. The microchip viscometer is economical and is shown to be accurate, which is very important for the simulation and control of lab-on-a-chip experiments.

Acoustothermal heating of polydimethylsiloxane microfluidic system

We report an observation of rapid (exceeding 2,000 K/s) heating of polydimethylsiloxane (PDMS), one of the most popular microchannel materials, under cyclic loadings at high (~MHz) frequencies. A microheater was developed based on the finding. The heating mechanism utilized vibration damping in PDMS induced by sound waves that were generated and precisely controlled using a conventional surface acoustic wave (SAW) microfluidic system. The refraction of SAW into the PDMS microchip, called the leaky SAW, takes a form of bulk wave and rapidly heats the microchannels in a volumetric manner. The penetration depths were measured to range from 210 μm to 1290 μm, enough to cover most sizes of microchannels. The energy conversion efficiency was SAW frequency-dependent and measured to be the highest at around 30 MHz. Independent actuation of each interdigital transducer (IDT) enabled independent manipulation of SAWs, permitting spatiotemporal control of temperature on the microchip. All the advantages of this microheater facilitated a two-step continuous flow polymerase chain reaction (CFPCR) to achieve the billion-fold amplification of a 134 bp DNA amplicon in less than 3 min.

Fast fabrication of microfluidic devices using a low-cost prototyping method

Conventional ways to produce microfluidic devices cost a lot due to the requirements for cleanroom environments and expensive equipment, which prevents the wider applications of microfluidics in academia and in industry. In this paper, a dry film photoresist was utilized in a simple way to reduce the fabrication cost of microfluidic masters. Thus, a fast prototyping and fabrication of microstructures in polydimethylsiloxane microchips through a replica molding technology was achieved in a low-cost setting within 2.5 h. Subsequently, major manufacturing conditions were optimized to acquire well-resolved microfluidic molds, and the replicated microchips were validated to be of good performance. A T-junction channel microchip was fabricated by using a dry film master to generate water droplets of uniform target size. Meanwhile, a gated injection of fluorescein sodium and a contactless conductivity detection of Na+ were both performed in a crosslink channel microchip via capillary electrophoresis, in other words, this fast prototyping and fabrication method would be an efficient, economical way to embody structural design into microfluidic chips for various applications.

Generation of Complex, Dynamic Temperature Gradients in a Disposable Microchip

We present a temperature gradient (TG) generation method using acoustothermal heating of polydimethylsiloxane (PDMS) by surface acoustic wave (SAW) microfluidic system. Independent and precise control of each SAW from a slanted interdigital transducer (IDT) enables a one-dimensional (1D) spatiotemporal control of temperature in a PDMS microchip. Various TGs were created in gas and liquid. The system creates switchable, dynamic and complex TGs in a disposable microchip.

Fluorometric flow-immunoassay for alkylphenol polyethoxylates on a microchip containing a fluorescence detector comprised of an organic light emitting diode and an organic photodiode

A compact fluorescence detector was constructed on a microchip from an organic light emitting diode (OLED) as the light source and an organic photodiode (OPD) as the photo-detector and was used in an immunoassay for alkylphenol polyethoxylates (APE). The OLED based on a terbium complex emitted a sharp light at the main wavelength of 546nm with a full width at half maximum of 9nm. The incident photo-to-current conversion efficiency (IPCE) of the OPD fabricated with Fullerene 70 (C70) and tris[4-(5-phenylthiopen-2-yl)phenyl]-amine (TPTPA) was approximately 44% for light at a wavelength of 586nm. The performance of the fluorescence detector was evaluated for the determination of resorufin (λem=586nm) and the photocurrent of the OPD due to the fluorescence of resorufin was proportional to the concentration of resorufin in the range from 0 to 18µM with a detection limit (S/N=3) of 0.6µM. The fluorescence detector was successfully utilized in a competitive enzyme-linked immunosorbent assay for APE, where an anti-APE antibody was immobilized on the surface of the channel of the Polydimethylsiloxane (PDMS) microchip or on the surface of magnetic microbeads. After an immunoreaction with a sample solution of APE containing a horse radish peroxidase (HRP)-labeled APE, the fluorescence of resorufin generated just after introduction of a mixed solution of Amplex Red and H2O2 was measured using the fluorescence detector. The calibration curve for the photocurrent signals of the OPD due to the fluorescence of resorufin against the logarithmic concentration of APE was sigmoidal in shape. The detection limits defined as IC80 were ca. 1ppb and ca. 2ppb, respectively, for the methods using the anti-APE antibody immobilized on the surface of the microchannel and in the case where the antibody was immobilized on the surface of magnetic microbeads.

Direct inkjet printing of micro-scale silver electrodes on polydimethylsiloxane (PDMS) microchip

Recently, direct inkjet printing of conductive solutions has received much attention in the microfluidics and lab-on-a-chip community because of its low-cost and mask-free deposition of electrodes on various substrates. However, the investigation of micro-scale direct inkjet printing on the polydimethylsiloxane (PDMS) substrate has not been completed. Here we present a direct inkjet printing technique to produce narrow (40–90 µm) silver microelectrodes on PDMS. Extensive experimental characterization studies on the pattern uniformity and electrical properties of the printed silver lines are presented. The effect of major printing parameters such as drop spacing, sintering temperature and duration, platen temperature, and nozzle temperature have been thoroughly investigated. We also investigated multiple layer printing as well as the effects of thermal expansion and mechanical bending. In order to demonstrate the utility of the inkjet-printed silver microelectrode, we fabricated both quadruple and castellated electrodes, and conducted dielectrophoretic manipulation of microbeads. The results clearly show that the printed silver electrodes can be used for electrokinetic applications in PDMS microchip devices. We believe that the direct inkjet printing of silver ink on PDMS presented here can provide a very convenient way of creating microelectrodes on PDMS devices for a variety of applications in the MEMS, microfluidics, and lab-on-a-chip communities.

Fabrication of a three-layer SU-8 mould with inverted T-shaped cavities based on a sacrificial photoresist layer technique.

A novel method for fabricating a three-layer SU-8 mould with inverted T-shaped cavities is presented. The first two SU-8 layers were spin coated and exposed separately, and simultaneously developed to fabricate the bottom and the horizontal part of the inverted T-shaped cavity. Then, a positive photoresist was filled into the cavity, and a wet lapping process was performed to remove the excess photoresist and make a temporary substrate. The third SU-8 layer was spin coated on the temporary substrate to make the vertical part of the inverted T-shaped cavity. The sacrificial photoresist layer can prevent the first two SU-8 layers from being secondly exposed, and make a temporary substrate for the third SU-8 layer at the same time. Moreover, the photoresist can be easily removed with the development of the third SU-8 layer. A polydimethylsiloxane (PDMS) microchip with arrays of T-shaped cantilevers for studying the mechanics of cells was fabricated by using the SU-8 mould.

A hydrodynamic microchip for formation of continuous cell chains

Here, we demonstrate the unique features of a hydrodynamic based microchip for creating continuous chains of model yeast cells. The system consists of a disk shaped microfluidic structure, containing narrow orifices that connect the main channel to an array of spoke channels. Negative pressure provided by a syringe pump draws fluid from the main channel through the narrow orifices. After cleaning process, a thin layer of water is left between the glass substrate and the polydimethylsiloxane microchip, enabling leakage beneath the channel walls. A mechanical clamp is used to adjust the operation of the microchip. Relaxing the clamp allows leakage of liquid beneath the walls in a controllable fashion, leading to formation of a long cell chain evenly distributed along the channel wall. The unique features of the microchip are demonstrated by creating long chains of yeast cells and model 15 μm polystyrene particles along the side wall and analysing the hydrogen peroxide induced death of patterned cells.

Interfacing microfluidic chip-based chromatography with flame atomic absorption spectrometry for the determination of chromium(VI)

This work demonstrates the first study on interfacing of microfluidic chip-based chromatography to flame atomic absorption spectrometry (FAAS) for a simple and fast separation or preconcentration and subsequent determination of species. The developed low-cost polydimethylsiloxane microchip includes 12 microcolumns of conventional C18 (5μm) chromatographic particles, making the parallel chromatographic separation/preconcentration and the direct introduction of the effluent into the spectrometer possible. In order to test this hyphenated microchip–FAAS system, the Cr(VI) was selected to be determined. The adsorbed Cr(VI) was eluted with methanol, and the 30μL effluents were collected into plastic vessels inserted into the ends of the microcolumns. Then, the effluents were analyzed by FAA spectrometer using micro-injection of the effluents as discrete samples. The separation/preconcentration and FAAS determination of 12 samples needed less than 5min due to the multiplex feature of the proposed system. The overall capacity of the microcolumn (C18, 5μm beads, 20mm×1mm×0.1mm) was calculated to be 0.45μg/mm for Cr(VI). Loading only 80μL samples onto the microchip and nebulizing the methanolic effluent of 30μL into the FAA spectrometer, 0.0031μg/mL limit of detection was obtained.

An integrated microfludic device for culturing and screening of Giardia lamblia

In vitro culturing of trophozoites was important for research of Giardia lamblia (G. lamblia), especially in discovery of anti-Giardia agents. The current culture methods mainly suffer from lab-intension or the obstacle in standardizing the gas condition. Thus, it could benefit from a more streamlined and integrated approach. Microfluidics offers a way to accomplish this goal. Here we presented an integrated microfluidic device for culturing and screening of G. lamblia. The device consisted of a polydimethylsiloxane (PDMS) microchip with an aerobic culture system. In the microchip, the functionality of integrated concentration gradient generator (CGG) with micro-scale cell culture enables dose–response experiment to be performed in a simple and reagent-saving way. The diffusion-based culture chambers allowed growing G. lamblia at the in vivo like environment. It notable that the highly air permeable material of parallel chambers maintain uniform anaerobic environment in different chambers easily. Using this device, G. lamblia were successfully cultured and stressed on-chip. In all cases, a dose-related inhibitory response was detected. The application of this device for these purposes represents the first step in developing a completely integrated microfluidic platform for high-throughput screening and might be expanded to other assays based on in vitro culture of G. lamblia with further tests.

Endothelial cells derived from embryonic stem cells respond to cues from topographical surface patterns

The generation of micro- and nano-topography similar to those found in the extra cellular matrix of three-dimensional tissues is one technique used to recapitulate the cell-tissue physiology found in the native tissues. Despite the fact that ample studies have been conducted on the physiological significance of endothelial cells alignment parallel to shear stress, as this is the normal physiologic arrangement for healthy arterial EC, very few studies have examined the use of topographical signals to initiate endothelial cell alignment. Here, we have examined the ability for our mouse embryonic stem cell-derived endothelial cells (ESC-EC) to align on various microchip topographical systems. Briefly, we generated metal molds with ‘wrinkled’ topography using 1) 15 nm and 2) 30 nm of gold coating on the pre-strained polystryene (PS) sheets. After thermal-induced shrinkage of the PS sheets, polydimethylsiloxane (PDMS) microchips were then generated from the wrinkled molds. Using similar Shrink™-based technology, 3) larger selectively crazed acetone-etched lines in the PS sheets, and 4) fully crazed acetone-treated PS sheets of stochastic topographical morphology were also generated. The 15 nm and 30 nm gold coating generated ‘wrinkles’ of uniaxial anisotropic channels at nano-scaled widths while the crazing generated micron-sized channels. The ESC-EC were able to respond and align on the 320 nm, 510 nm, and the acetone-etched 10.5 μm channels, but not on the fully ‘crazed’ topographies. Moreover, the ESC-EC aligned most robustly on the wrinkles, and preferentially to ridge edges on the 10.5 μm-sized channels. The ability to robustly align EC on topographical surfaces enables a variety of controlled physiological studies of EC-EC and EC-ECM contact guidance, as well as having potential applications for the rapid endothelialization of stents and vascular grafts.

A droplet-based novel approach for viable and low volume consumption surface plasmon resonance bio-sensing inside a polydimethylsiloxane microchip

Over the course of last two decades, surface plasmon resonance (SPR) has emerged as a viable candidate for label-free detection and characterization for a large pool of biological interactions, ranging from hybridization of oligonucleotides to high throughput drug-screening. Conventional SPR bio-sensing involves a step-response method where the SPR sensorgram in response to a switched sequential flow of analyte and buffer is plotted in real-time and fitted to an exponential curve to extract the associative and dissociative reaction rates. Such measurement schemes involve continuous flow conditions where a substantial reagent volume is consumed and is subject to dispersive mixing at flow switching zones. In this paper, we demonstrate a new plug-train SPR technique in a microfluidic chip that separates and singulates solvent plugs in analyte and buffer by an immiscible air phase. Bio-samples are first discretized within plug droplets with volumes in order of few hundred nanoliters or less followed by pressure-driven transport onto SPR sensing sites of this hydrophobically modified SPR microdevise. The kinetic constants ka and kd for a model protein-small molecule interaction pair are extracted from a plug-train signal and are shown to be in reasonable agreement with our previous reports.

Modeling of capacitively coupled contactless conductivity detection on microfluidic chips

A novel equivalent circuit model of capacitively coupled contactless conductivity detection (C4D) on microfluidic chips is presented. The impedance of the solution in microchannels facing the two electrodes for C4D was first introduced in the model of C4D on microfluidic chips. The electrodes and the solution facing electrodes were divided into individual segments in the model, and the effect of the length of divided segments on the model was studied. A back-calculating method was put forward to calculate the stray capacitance between the electrodes, and the variation between the calculated value and the simulated value was only 6 %. To evaluate the accuracy of the model, a hybrid poly (methyl methacrylate) (PMMA)/polydimethylsiloxane (PDMS) microchip was fabricated and a simple model was built. Compared with the outputs of the simple model, the data predicted by the novel model show a much closer fit to experimental results, and the variations were within 8 % over a wide concentration range of 1–500 μm for potassium chloride.

A novel open-tubular capillary electrochromatography using β-cyclodextrin functionalized graphene oxide-magnetic nanocomposites as tunable stationary phase

Chip-based enantioselective open-tubular capillary electrochromatography (OT-CEC) with β-cyclodextrin (β-CD) conjugated graphene oxide-magnetic nanocomposites (GO/Fe3O4 NCs) as stationary phase was developed. GO/Fe3O4 NCs with high magnetic responsivity, excellent solubility and high dispersibility in water were prepared through a facile and controllable in situ chemical deposition strategy. β-CD was then adsorbed onto the GO/Fe3O4 surface to form GO/Fe3O4/β-CD NCs which were localized to the pre-nominated position in polydimethylsiloxane (PDMS) microchannels with the help of magnets. The resultant GO/Fe3O4/β-CD NCs not only have the magnetism of Fe3O4 NPs that make them easily manipulated by an external magnetic field, but also have the larger surface which can incorporate much more chiral selector molecules. In addition, the successful β-CD decorations endowed GO/Fe3O4/β-CD NCs with excellent wettability and led to enhanced stability against high ionic strength. Compared with the native PDMS microchip, the modified surfaces exhibited more stable and suppressed electroosmotic mobility, and less nonspecific adsorption toward analytes. Successful baseline separation of tryptophan enantiomers was achieved in less than 50s with a resolution factor of 1.65 utilizing a separation length of 37mm coupled with in-column amperometric detection. Factors that influence the chiral separation resolution were examined. Under the optimized conditions, the proposed modified chip revealed adequate repeatability concerning run-to-run and day-to-day. These results show that the use of GO/Fe3O4/β-CD NCs within microfluidic channels hold great promise for a variety of analytical schemes.

Selective modification for polydimethylsiloxane chip by micro-plasma

We report a method to selectively modify polydimethylsiloxane (PDMS) chip in a fast and facile way using micro-plasma approach in the atmospheric-pressure. Pure He and He/acrylic acid plasma were ignited directly in different channels of PDMS microchip. Our experiments results yielded strong hydrophilic property on the surface of PDMS by the plasma treatment.

Particle analysis and differentiation using a photovoltaic cell

A method is proposed for the sizing and counting of fluorescent and non-fluorescent particles of various sizes on a poly-dimethylsiloxane microchip. In the proposed approach, the detection region of the microchip is illuminated by a laser, which is then incident on a power-free photovoltaic cell. As the particles (both fluorescent and non-fluorescent) pass through the detection region, they block the laser beam, causing a reduction in the output voltage of the cell. The voltage signal is interfaced to a PC and is used to determine both the size and the number of the particles. Meanwhile, the fluorescence signal generated by the fluorescent particles within the sample is detected by an avalanche photodetector and is used to differentiate between the fluorescent and non-fluorescent particles in the sample. The effectiveness of the proposed approach is demonstrated using fluorescent-labeled beads with means diameters of 5, 8 and 10 µm, respectively, and unlabeled beads with a mean diameter of 7.2 µm. The experimental results confirm that the forward scattered light signal generated by the photovoltaic cell enables both the size and the number of the particles to be reliably determined. Moreover, it is shown that the number of non-fluorescent particles within the sample can be easily determined by comparing the signals received from the photovoltaic cell and avalanche photodetector, respectively.

Separation and Determination of Three Phenolic Xenoestrogens in Industrial Wastewater by Micellar Electrokinetic Chromatography on Polydimethylsiloxane Microchip

The separation on microchip provides the advantages including high efficiency, increased throughput, reduced quantities of hazardous materials, cost saving, relatively facile instrumentation, improved portability, etc. A technique of micellar electrokinetic chromatography (MEKC) coupled with amperometric detection has been actualized on a polydimethylsiloxane microchip for the rapid separation and determination of three phenolic xenoestrogens as octylphenol (OP), 4-nonylphenol (4-NP), and bisphenol A (BPA). The baseline separation of these phenolic xenoestrogens is successfully obtained within 55 s under the optimized MEKC conditions with borate running buffer of pH 8.0 containing sodium dodecyl sulfate and β-cyclodextrin. The linear range for OP, 4-NP, and BPA are 20–1,000, 15–1,000, and 20–1,000 μg/L with the detection limit of 5.0, 4.0, and 3.0 μg/L, respectively. The present method is successfully applied for the determination of these phenolic xenoestrogens in some industrial wastewater samples from mainland of China with the recoveries ranged from 90.2 to 109.4 %.