Poly Methyl Methacrylate Chip Research Articles

In–situ self–reduction preparation of Ti3C2Tx/Ag on flexible PMMA chip for quantitative detection of SARS–CoV–2

Surface enhanced Raman scattering (SERS) has proven to be of great superiority in the assay and prevention of infectious disease attributed to its rapid and specific fingerprint recognition ability toward trace molecules. However, it is still crucial to develop portable and precise chip for the reliable realization of on–site and large–scale screening. Here, a robust in–situ reduction strategy was employed to prepare Ti3C2Tx@Ag nanocomposites, which was installed on a polymethyl methacrylate (PMMA) matrix to construct a novel flexible SERS chip with self–rectification capability. The resulting Ti3C2Tx@Ag nanocomposites exhibited both electromagnetic and chemical enhancement effects, enabling high SERS activity. In particular, the as–developed SERS chip demonstrated fascinating signal reproducibility and quantitative detection capability by conducting the intrinsic Raman signal of PMMA as an internal standard. Furthermore, the PMMA–Ti3C2Tx@Ag chip facilitated the visualization of severe acute respiratory syndrome coronavirus 2 (SARS–CoV–2) nucleocapsid (N) with ultra–low limit of detection, stressing the potential application of this smart self–rectification SERS chip with high activity in real time and rapid monitoring of sudden infectious diseases.

Microfluidic-based ion-selective thermoplastic electrode array for point-of-care detection of potassium and sodium ions.

A microfluidic paper-based thermoplastic electrode (TPE) array has been developed for point-of-care detection of Na+ and K+ ions using a custom-made portable potentiometer. TPEs were fabricated using polystyrene as the binder and two different types of graphite to compare theelectrode performance. The newly designed TPE array embedded in a polymethyl methacrylate chip consists of two working electrodes modified with carbon black nanomaterial and an ion-selective membrane, and an all-solid-state reference electrode modified with Ag/AgCl ink and poly(butyl methacrylate-co-methyl methacrylate) membrane via drop-casting. Ion-selective membrane compositions and conditioning steps were optimized. Under optimized conditions, ion-selective TPEs demonstrated fast response time (4s) and good stability. The TPE array demonstrated a Nernstian behavior for K+ with a sensitivity of 59.2 ± 0.2mV decade-1 and near-Nernstian response for Na+ with a sensitivity of 54.0 ± 1.1mV decade-1 in the range 10-1 - 10-4M and 1 - 10-3M, respectively. The detection limits were 1 × 10-5M and 1 × 10-4M for K+ and Na+, respectively. In addition, a K+ and Na+ selective microfluidic paper-based analytical device (µPAD) was applied to artificial serum analysis and found in good agreement with average recoveries of 101.3% and 99.7%, respectively, suggesting that the developed ISE array is suitable for detection of sodium and potassium in complex matrix.

Gel Electrophoresis Chip Using Joule Heat Self-Dissipation, Short Run Time, and Online Dynamic Imaging.

Gel electrophoresis (GE) is one of the most general tools in biomedicine. However, it suffers from low resolution, and its mechanism has not been fully revealed yet. Herein, we presented the dispersion model of w2 (t) ∝ Tt, showing the band dispersion (w) via temperature (T) and running time (t) control. Second, we designed an efficient GE chip via the time control and rapid Joule heat self-dissipation by thermal conductive plastic (TCP) and electrode buffer. Third, we conducted the simulations on TCP and polymethylmethacrylate (PMMA) chips, unveiling that (i) the temperature of TCP was lower than the PMMA one, (ii) the temperature uniformity of TCP was better than the PMMA one, and (iii) the resolution of TCP was superior to the PMMA one. Fourth, we designed both TCP and PMMA chips for experimentally validating the dispersion model, TCP chip, and simulations. Finally, we applied the TCP chip to thalassemia and model urine protein assays. The TCP chip has merits of high resolution, rapid run of 6-10 min, and low cost. This work paves the way for greatly improving electrophoretic techniques in gel, chip, and capillary via temperature and time control for biologic study, biopharma quality control, clinical diagnosis, and so on.

Collateral flow at circle of Willis in healthy condition.

Experimental simulation of cerebrovascular system would be very beneficial tool to evaluate millions of human body cascade sequence. The Circle of Willis (CoW) recently named Cerebral Arterial Circle (CAC) is a main loop structure of cerebral circulatory system which positioned at the cranium base. In this research, we investigate cerebral artery flow pattern in cerebral arteries including afferent, Willisian, and efferent arteries of CAC emphasizing on communicating and connecting arteries which are main routes in CAC and as a risky sites when autoregulation is occurred in terminal parts of middle cerebral arteries (MCAs) by PMMA (Polymethyl methacrylate) chip and high quality camera which depict Sequential images. This novelty study analyze flow pattern in CAC that have been challenging subject area for many years which have investigated by scientists yet, because flow pattern in CAC indicate complication progression. This research tries to construct new platform in cerebral circulation analyzing method by reliable experimental in-vitro approach. The outcomes of this study demonstrate that communicating arteries especially anterior communicating artery (ACoA) is main artery in CAC flow distribution.

Fused Deposition Modeling of Microfluidic Chips in Polymethylmethacrylate.

Polymethylmethacrylate (PMMA) is one of the most important thermoplastic materials and is a widely used material in microfluidics. However, PMMA is usually structured using industrial scale replication processes, such as hot embossing or injection molding, not compatible with rapid prototyping. In this work, we demonstrate that microfluidic chips made from PMMA can be 3D printed using fused deposition modeling (FDM). We demonstrate that using FDM microfluidic chips with a minimum channel cross-section of ~300 µm can be printed and a variety of different channel geometries and mixer structures are shown. The optical transparency of the chips is shown to be significantly enhanced by printing onto commercial PMMA substrates. The use of such commercial PMMA substrates also enables the integration of PMMA microstructures into the printed chips, by first generating a microstructure on the PMMA substrates, and subsequently printing the PMMA chip around the microstructure. We further demonstrate that protein patterns can be generated within previously printed microfluidic chips by employing a method of photobleaching. The FDM printing of microfluidic chips in PMMA allows the use of one of microfluidics’ most used industrial materials on the laboratory scale and thus significantly simplifies the transfer from results gained in the lab to an industrial product.

Study of the Effect of Material of Microfluid Chip on the Polymerase Chain Reaction

Effect of several polymer materials from different manufacturers on the polymerase chain reaction is tested. Absence of adsorption of reaction components at room temperature is demonstrated for a polymethylmethacrylate chip. However, a decrease in the reaction efficiency at low DNA concentrations is observed for the polymerase chain reaction in the microchip.

End-point dual specific detection of nucleic acids using CRISPR/Cas12a based portable biosensor

A CRISPR/Cas12a based portable biosensor (Cas12a-PB) was developed to simultaneously visually detect CaMV35S promoter and Lectin gene from genetically modified (GM) soybean powders (Roundup Ready@). The Cas12a-PB, mainly made of polymethylmethacrylate (PMMA) and PMMA tape, has a connection structure, three channels and three detection chambers. The CRISPR/Cas12a detection reagents were preloaded in detection chambers and the reaction tube was connected to the connection structure by screw threads. After amplification, the amplicons were gone into three detection chambers by swinging the Cas12a-PB to conduct dual detection. Positive samples would produce green fluorescence while negative samples were black under the irradiation of 490 nm LED light. In this study, the Cas12a-PB successively combined with ordinary PCR, rapid PCR and loop-mediated isothermal amplification (LAMP) to achieve dual detection, which made detection process more convenient and portable. As low as 0.1% transgenic ingredients in soybean powders could be detected and the specificity of Cas12a-PB was confirmed with GM maize powders (MON810, GA21), GM soybean powders (DP305423), non-GM peanut and rice as targets. In the end, an amplification chamber combining with Cas12a-PB on a PMMA chip was further designed to eliminate the use of reaction tube and mineral oil, which made operation simpler. The established Cas12a-PB would provide a new reliable solution for multiple targets detection in clinic diagnostics, food safety, etc.

Fabrication of Rugged and Reliable Fluidic Chips for Autonomous Environmental Analyzers Using Combined Thermal and Pressure Bonding of Polymethyl Methacrylate Layers.

The fabrication of highly reliable and rugged fluidic chips designed for use in autonomous analyses for nutrient monitoring is described. The chips are based on a two-layer configuration with the fluidic channels produced in one layer using precision micromilling. The second capping layer contains through holes for sample/standard and reagent addition and waste removal post-analysis. Two optically clear polymethyl methacrylate (PMMA) windows are integrated into the opaque PMMA chip, orthogonal to a 22.5 mm-long section of the channel downstream from a serpentine reagent and sample/standard mixing region. An LED source is coupled into the channel through one of the windows, and the light intensity is monitored with a photodiode located at the distal end of the channel outside the second optically clear window. Efficient coupling of the source through the channel to the detector is achieved using custom-designed alignment units produced using 3D printing. In contrast to fluidic chips produced using solvent adhesion, the thermal-/pressure-bonded simplified method presented removes the need for surface treatment. Optimization of the thermal/pressure conditions leads to very strong adhesion between the PMMA layers, requiring forces in the region of 2000 N to separate them, which is necessary for the use in long-term deployments. Profilometry imaging shows minimal evidence of channel distortion after bonding. Finally, we show the potential of these techniques for environmental applications. The fluidic chips were integrated into prototype nutrient analyzers that display no evidence of leakage in extensive lab tests involving 2500 phosphate measurements using the yellow (vanadomolybdophosphoric acid) method. Similarly, excellent analytical performance (LOD is 0.09 μM) is reported for a 28-day field trial comprising 188 in situ autonomous phosphate measurements (564 measurements) in total including calibration.

Systematic time-dependent visualization and quantitation of the neurogenic rate in brain organoids

Organoids mimicking the formation of the brain cortex have been demonstrated to be powerful tools for developmental studies as well as pathological investigations of brain malformations. Here, we report an integrated approach for the quantification of temporal neural production (neurogenic rate) in organoids derived from embryonic brains. Spherical tissue fragments with polarized cytoarchitectures were incubated in multiple cavities arranged in a polymethylmethacrylate chip. The time-dependent neurogenic rate in the organoids was monitored by the level of EGFP under the promoter of Tbr2, a transcription factor that is transiently expressed in neural fate-committed progenitors during corticogenesis. Importantly, our monitoring system exhibited a quick response to DAPT, a drug that promotes neural differentiation. Furthermore, we successfully quantified the temporal neurogenic rate in a large number of organoids by applying image processing that semi-automatically recognized the positions of organoids and measured their signal intensities from sequential images. Taken together, we provide a strategy to quantitate the neurogenic rate in brain organoids in a time-dependent manner, which will also be a potent method for monitoring organoid formation and drug activity in other tissue types.

Flow-Injection Spectrophotometric Determination of Cysteine in Biologically Active Dietary Supplements

A procedure is developed for the flow-injection spectrophotometric determination of cysteine in dietary supplements based on the formation of a reduced phosphomolybdic complex by the analyte in mixing a sample solution with a solution of ammonium 18-molybdophosphate. For the flow-injection determination of cysteine, a polymethyl methacrylate chip is fabricated, in the channels of which the analytical form was obtained and detected. The detection limit for cysteine is 3 × 10–6 M and the throughput is 240 determinations per hour. The procedure was tested in the analysis of different samples of dietary supplements; the results obtained were verified by HPLC.

A Novel Fabrication Procedure for the Preparation of Polymethyl Methacrylate Lab-on-a-Chip Substrates

Modification of a minicomputer numerical controller with a low cost diode laser for fabrication of polymethyl methacrylate chips has been demonstrated. The maximum power of the diode laser was 5 W at 808 nm. The scanning speed was 1.0–10.0 mm s−1 by the movement of x and y stages. The patterns of microchannels on the chip were designed using drawing software and then applied to software that controlled the operation of the controller. The parameters that affected the channel depth, width, and smoothness were studied by varying the laser power and speed of polymethyl methacrylate sheets with surface area of 4.0 × 6.0 cm2 and thickness of 0.1 cm. The optimum conditions were used to fabricate microchannels on each sheet giving different depths and widths over the range of 74 µm to 554 µm and 147 µm to 393 µm, respectively. The most appropriate conditions for polymethyl methacrylate chip fabrication were 5 W and 3 mm s−1 for the laser diode power and the speed, respectively. The optimal dimensions of the microchannel on the polymethyl methacrylate sheet were 226 µm in width and 202 µm in depth based on sensitivity, reproducibility, and low background signals. The microchannels were sealed with a polymethyl methacrylate cover plate by thermal bonding. The resulting chips were tested for iron(III) determination in water based on microreverse flow analysis.

UV 개질된 PMMA 미세유체 장치의 열가소성 폴리머 용융 접합

Thermoplastic fusion bonding is widely used to seal polymer microfluidic devices and optimal bonding protocol is required to obtain a successful bonding, strong bonding force without channel deformation. Besides, UV modification of the PMMA (poly-methyl methacrylate) is commonly used for chemical or biological application before the bonding process. However, study of thermal bonding for the UV modified PMMA was not reported yet. Unlike pristine PMMA, the optimal bonding parameters of the UV modified PMMA were 103˚C, 71 kPa, and 35 minutes. A very low aspect ratio micro channel (AR=1:100, 20 μm depth and 2000 μm width) was successfully bonded (over 95%, n>100). Moreover, thermal bonding of multi stack PMMA chips was successfully demonstrated in this study. The results may applicable to fabricate a complex 3 dimensional microchannel networks.

Surface characteristics and electrical properties of PMMA chips for incubation-type planar-patch-clamp biosensors

Polymethylmethacrylate (PMMA) plates were successfully applied as sensor chips in an incubation-type planar patch clamp (IPPC). Hot embossing both sides formed the PMMA plates, and a focused ion beam realized micropores. The low seal resistance of the IPPC was investigated by analyzing the surface roughness of the chips. Atomic force microscopy (AFM) showed that the chip surface had a roughness of several nanometers due to the molding process. Coating the molded surface with an anti-adhesive compound further increased the surface roughness of the PMMA chip because the anti-adhesive compound itself had a large roughness and in some case, the compound partially peeled off while detaching the mold. Similarly, coating a chip with extracellular matrix (ECM) poly-l-lysine (PLL) also increased the surface roughness. The measured seal resistance of the PMMA chip for an HEK293 cell was in the range of 4–15MΩ. The low seal resistance was attributed to the sharp-edge structure of the micropore and the surface roughness of the chip. Nevertheless, the whole cell current was successfully recorded from HEK293 cells expressing channel rhodopsin wide receiver (ChRWR) using salt-bridge-type stable Ag/AgCl electrodes. Another advantage of the PMMA sensor chip was the small parasitic capacitance.

Fully Automated On-Chip Imaging Flow Cytometry System with Disposable Contamination-Free Plastic Re-Cultivation Chip

We have developed a novel imaging cytometry system using a poly(methyl methacrylate (PMMA)) based microfluidic chip. The system was contamination-free, because sample suspensions contacted only with a flammable PMMA chip and no other component of the system. The transparency and low-fluorescence of PMMA was suitable for microscopic imaging of cells flowing through microchannels on the chip. Sample particles flowing through microchannels on the chip were discriminated by an image-recognition unit with a high-speed camera in real time at the rate of 200 event/s, e.g., microparticles 2.5 μm and 3.0 μm in diameter were differentiated with an error rate of less than 2%. Desired cells were separated automatically from other cells by electrophoretic or dielectrophoretic force one by one with a separation efficiency of 90%. Cells in suspension with fluorescent dye were separated using the same kind of microfluidic chip. Sample of 5 μL with 1 × 106 particle/mL was processed within 40 min. Separated cells could be cultured on the microfluidic chip without contamination. The whole operation of sample handling was automated using 3D micropipetting system. These results showed that the novel imaging flow cytometry system is practically applicable for biological research and clinical diagnostics.

Fabrication of plastic microchips with gold microelectrodes using techniques of sacrificed substrate and thermally activated solvent bonding

A novel fabrication process of polymethyl methacrylate (PMMA) microfluidic chip with embedded gold microelectrodes has been developed. The PMMA chips consist of a microfluidic part, which is made by micro-milling and a mate with embedded gold electrodes made by sacrificed substrate technique. The developed sacrificed substrate technique combines photolithography, gold electrodeposition and embedding of the gold structures into a UV curable resin. Both microfluidic and electrode parts are bonded together by thermally activated solvent bonding. Isopropyl alcohol (IPA) is used as a bonding agent. Using the IPA solvent brings several advantages compared to conventional thermal bonding: (i) decreasing the bonding temperature, (ii) higher bond strength. Decreasing the bonding temperature is crucial for the whole process, because of prevention of electrode cracking, microfluidic channel deformation and clogging. Higher bond strength prevents a liquid leakage and generally improves the microdevice durability. The fabrication process was successfully tested in construction of a four-electrode AC micro-conductometer.

Use of a heterogeneous buffer combination in microchip electrophoresis for high‐resolution separation by on‐line concentration of DNA samples

We have developed a novel high-resolution separation technique of DNA fragments in a heterogeneous combination of a sample buffer and a separation buffer. The use of a heterogeneous buffer combination is a simple method for on-line concentration of DNA fragments, in which a sample buffer is simply exchanged with one including taurine anions. The mobility of taurine anions, co-ions for DNA, is lower than the that of acetate anions in a separation buffer. The difference in the mobility invokes transient isotachophoresis. The current technique allows DNA fragments to be effectively concentrated and the separation length of microchips to be shorter than that of conventional ones by a factor of three without deterioration in separation resolution and any modification of a chip design. Fragments of 100-bp DNA ladders (100-1000 bp) were separated with high resolution (0.72-10.7) within 60 s with a 10 mm separation length on a polymethyl methacrylate chip. Furthermore, fragments of 10-bp DNA ladders (10-330 bp) were separated with high resolution (0.69-2.00) with a 10 mm separation length within 50 s without band broadening. The current achievements will make it possible to fabricate compact devices for microchip electrophoresis.

Optical PMMA Chip Suitable for Multianalyte Detection

A new optical platform for the interrogation of a polymethylmethacrylate (PMMA) multichannel array for chemical and biochemical parameters is described. It consists of a plastic chip formed by two pieces of PMMA properly shaped in order to obtain different microchannels, 500 mum in width and 400 mum in height. A fluorescent sensing layer is immobilized on the internal wall of the microchannel. The emitted light travels along the thickness of the upper PMMA piece and it is detected with an optical fiber connected with a spectrum analyzer. The anisotropy of the fluorescence emitted by the fluorophore immobilized at the liquid/plastic interface gives rise to preferential directions of the emitted fluorescence. The collection of the fluorescence at an angle of 50deg implies an amplification of roughly one order of magnitude in the ratio between the fluorescence signal and the scattered light. The optical platform was further characterized as a pH sensor and as potential chip for immunoassay. In the first case, the fluorescein dye is directly immobilized onto the internal wall of the channel and the fluorescence changes are measured as a function of the pH of the flowing sample. In the second case, a direct antigen-antibody interaction is carried out. The mouse-IgG is covalently immobilized onto the internal wall of the channel and the Cy5-labeled anti-mouse IgG is used for the specific interaction.

Microfluidic chip made of COP (cyclo-olefin polymer) and comparion to PMMA (polymethylmethacrylate) microfluidic chip

Hot embossing method was studied for the fabrication of microfluidic microchannels on COP (cyclo-olefin polymer) chip. The ending temperature of heating stage, i.e. embossing reference temperature ( T r), was determined by the viscoelastic property of the COP, according to variable temperature quasi-creep experiments. Taguchi method was used to decide the processing parameters in cooling and demolding stage. The optimized parameters of hot embossing are: embossing reference temperature 143 °C, temperature and pressure holding time 2 min, pressure at cooling and demolding stage 1.6 MPa, the demolding temperature in the lower heating plate 80 °C and temperature difference between the upper and lower heating plate 10 °C. Experiment shows that microchannel fabricated with the above parameters has high repeatability and low substrate deformation, average repeatability is 97.6% in width and 94.3% in depth. Experiments on background fluorescence, electrophoresis, and DNA separation and detection were carried out on COP and PMMA (polymethylmethacrylate) chips, respectively. Compared to the PMMA chip, the COP chip has higher signal to noise ratio, higher electrophoresis efficiency, and lower peak area relative standard deviation (R.S.D.). In separation 6 μg/ml ϕ X174 RF DNA Hinc II, 11 DNA fragments have been identified in less than 2 min on the COP chip, thus it has large application potential in biochemical analysis.

A cleaner and simple spectrophotometric micro-fluidic procedure for copper determination using nitroso-R salt as chromogenic agent

A cleaner and simple spectrophotometric method using microflow analysis (μFA) was performed. It consisted of a T-junction with microcoil on a polymethyl methacrylate (PMMA) chip which was fabricated by laser ablation and a molded polydimethylsiloxane (PDMS) as top plate. The fabricated PMMA chip was integrated with light emitting diode (LED) as light source and spectrometer as detector. The proposed device was applied to determining copper in water samples using nitroso-R salt as chromogenic reagent at 495 nm. It was found that the proposed μFA system was with less reagents and samples consumption with tiny waste generation. The relative standard deviation (R.S.D.) was less than 2% ( n = 11) with the percentage recovery of 98.0 ± 1.7% ( n = 7). The linear range for determination of copper in water samples was over the range of 0.05–3.0 μg mL −1 with a correlation coefficient ( r 2) of 0.999. The limit of detection (3 σ) was 47 ng mL −1 with a sample throughput of 30 h −1.

Applying Taguchi methods for solvent-assisted PMMA bonding technique for static and dynamic μ-TAS devices

This work examines numerous significant process parameters in the solvent-assistant Polymethyl methacrylate (PMMA) bonding scheme and presents two Micro-total-analysis System (micro-TAS) devices generated by adopting the optimal bonding parameters. The process parameters considered were heating temperature, applied loading, duration and solution. The effects of selected process parameters on bonding dimensions loss and strength, and subsequent optimal setting of the parameters were accomplished using Taguchi's scheme. Additionally, two micro-TAS devices were realized using a static paraffin microvalve and a dynamic diffuser micropump. The PMMA chips were carved using a CO2 laser that patterned device microchannels and microchambers. The operation principles, fabrication processes and experimental performance of the devices are discussed. This bonding technique has numerous benefits, including high bonding strength (240 kgf/cm2) and low dimension loss (2-6%). For comparison, this work also demonstrates that the normal stress of this technology is 2-15 times greater than that of other bonding technologies, including hot embossing, anodic bonding, direct bonding and thermal fusion bonding.