Simulation and fabrication of in vitro microfluidic microelectrode array chip for patterned culture and electrophysiological detection of neurons

To enable the detection and modulation of modularized neural networks in vitro,this study proposes a microfluidic microelectrode array chip for the cultivation,compartmentalization,and control of neural cells.The chip was designed based on the specific structure of neurons and the requirements for detection and modulation.Finite-element analysis of the chip’s flow field was conducted using the COMSOL Multiphysics software,and the simulation results show that the liquid within the chip can flow smoothly,ensuring stable flow fields that facilitate the uniform growth of neurons within the microfluidic channels.By employing MEMS technology in combination with nanomaterial modification techniques,the microfluidic microelectrode array chip was fabricated successfully.Primary hippocampal neurons were cultured on the chip,forming a well-defined neural network.Spontaneous electrical activity of the detected neurons was recorded,exhibiting a 23.7%increase in amplitude compared to neuronal discharges detected on an open-field microelectrode array.This study provides a platform for the precise detection and modulation of patterned neuronal growth in vitro,potentially serving as a novel tool in neuroscience research.

A novel integrated microfluidic chip for on-demand electrostatic droplet charging and sorting

On-demand droplet sorting is extensively applied for the efficient manipulation and genome-wide analysis of individual cells.However,state-of-the-art microfluidic chips for droplet sorting still suffer from low sorting speeds,sample loss,and labor-intensive preparation procedures.Here,we demonstrate the development of a novel microfluidic chip that integrates droplet generation,on-demand electrostatic droplet charging,and high-throughput sorting.The charging electrode is a copper wire buried above the nozzle of the microchannel,and the deflecting electrode is the phosphate buffered saline in the microchannel,which greatly simplifies the structure and fabrication process of the chip.Moreover,this chip is capable of high-frequency droplet generation and sorting,with a frequency of 11.757 kHz in the drop state.The chip completes the selective charging process via electrostatic induction during droplet generation.On-demand charged microdroplets can arbitrarilymove to specific exit channels in a three-dimensional(3D)-deflected electric field,which can be controlled according to user requirements,and the flux of droplet deflection is thereby significantly enhanced.Furthermore,a lossless modification strategy is presented to improve the accuracy of droplet deflection or harvest rate from 97.49% to 99.38% by monitoring the frequency of droplet generation in real time and feeding it back to the charging signal.This chip has great potential for quantitative processing and analysis of single cells for elucidating cell-to-cell variations.

Rapid fabrication of modular 3D paper-basedmicrofluidic chips using projection-based 3D printing

Paper-based microchips have different advantages,such as better biocompatibility,simple production,and easy handling,making them promising candidates for clinical diagnosis and other fields.This study describes amethod developed to fabricate modular three-dimensional(3D)paper-based microfluidic chips based on projection-based 3D printing(PBP)technology.A series of two-dimensional(2D)paper-based microfluidic modules was designed and fabricated.After evaluating the effect of exposure time on the accuracy of the flow channel,the resolution of this channel was experimentally analyzed.Furthermore,several 3D paper-based microfluidic chips were assembled based on the 2D ones using different methods,with good channel connectivity.Scaffold-based 2D and hydrogel-based 3D cell culture systems based on 3D paper-based microfluidic chips were verified to be feasible.Furthermore,by combining extrusion 3D bioprinting technology and the proposed 3D paper-based microfluidic chips,multiorgan microfluidic chips were established by directly printing 3D hydrogel structures on 3D paperbased microfluidic chips,confirming that the prepared modular 3D paper-based microfluidic chip is potentially applicable in various biomedical applications.

Droplet microfluidic chip for precise monitoring of dynamic solution changes

In this work,an automated microfluidic chip that uses negative pressure to sample and analyze solutions with high temporal resolution was developed.The chip has a T-shaped channel for mixing the sample with a fluorescent indicator,a flow-focusing channel for generating droplets in oil,and a long storage channel for incubating and detecting the droplets.By monitoring the fluorescence intensity of the droplets,the device could detect changes in solution accurately over time.The chip can generate droplets at frequencies of up to 42 Hz with a mixing ratio of 1:1 and a temporal resolution of 3–6 s.It had excellent linearity in detecting fluorescein solution in the concentration range 1–5μM.This droplet microfluidic chip provides several advantages over traditional methods,including high temporal resolution,stable droplet generation,and faster flow rates.This approach could be applied to monitoring calcium ions with a dynamic range from 102 to 107 nM and a detection limit of 10 nM.

In vitro study of emodin-induced nephrotoxicity in human renal glomerular endothelial cells on a microfluidic chip

Emodin is an effective component of rhubarb with positive pharmacological effects on human health.However,it is also toxic to different cells or tissues to varying degrees.The effects of emodin on glomerular endothelial cells(GECs)remain to be tested,and the documented works were always performed in vitro and hardly reflect the real physiological situation.To study the effects of emodin on GECs in a biomimetic environment,we utilized a microfluidic chip to assess the physiological reaction of human renal glomerular endothelial cells to various concentrations of emodin in this work.The results showed that emodin caused cytotoxicity,impaired glomerular filtration barrier integrity to macromolecules,and increased barrier permeability in a dose-dependent manner.With the increase in emodin concentration,the concentration of the pro-inflammatory cytokine tumor necrosis factor-α,interleukin(IL)-6,transforming growth factor-β1,and monocyte chemoattractant protein(MCP-1)increased while the production of inflammatory cytokine IL-6 first increased and then decreased with the increase in emodin concentration.Our findings shed new light on emodin-induced nephrotoxicity and provide insights for the application of microfluidic chip devices to reveal drug-cell interactions.

Advances in microfluidic chips based on islet hormone-sensing techniques

Diabetes mellitus is a global health problem resulting from islet dysfunction or insulin resistance.The mechanisms of islet dysfunction are still under investigation.Islet hormone secretion is the main function of islets,and serves an important role in the homeostasis of blood glucose.Elucidating the detailed mechanism of islet hormone secretome distortion can provide clues for the treatment of diabetes.Therefore,it is crucial to develop accurate,real-time,laborsaving,high-throughput,automated,and cost-effective techniques for the sensing of islet secretome.Microfluidic chips,an elegant platform that combines biology,engineering,computer science,and biomaterials,have attracted tremendous interest from scientists in the field of diabetes worldwide.These tiny devices are miniatures of traditional experimental systems with more advantages of timesaving,reagent-minimization,automation,high-throughput,and online detection.These features of microfluidic chips meet the demands of islet secretome analysis and a variety of chips have been designed in the past 20 years.In this review,we present a brief introduction of microfluidic chips,and three microfluidic chipsbased islet hormone sensing techniques.We focus mainly on the theory of these techniques,and provide detailed examples based on these theories with the hope of providing some insights into the design of future chips or whole detection systems.

Application of microfluidic chip technology in pharmaceutical analysis:A review

The development of pharmaceutical analytical methods represents one of the most significant aspects of drug development. Recent advances in microfabrication and microfluidics could provide new approaches for drug analysis, including drug screening, active testing and the study of metabolism. Microfluidic chip technologies, such as lab-on-a-chip technology, three-dimensional (3D) cell culture, organs-on-chip and droplet techniques, have all been developed rapidly. Microfluidic chips coupled with various kinds of detection techniques are suitable for the high-throughput screening, detection and mechanistic study of drugs. This review highlights the latest (2010–2018) microfluidic technology for drug analysis and discusses the potential future development in this field.

EXPERIMENTAL STUDY ON THE HOT EMBOSSING POLYMER MICROFLUIDIC CHIP

Experiments are used to study the fabrication of polymer microfluidic chip with hot embossing method. The pattern fidelity with respect to the process parameters is analyzed. Experiment results show that the relationship between the imprint temperature and the microchannel width is approximately exponential. However, the depth of micro channel isn＇t sensitive to the imprint temperature. When the imprint pressure is larger than 1 MPa and the imprint time is longer than 2 min, the increasing of imprint pressure and holding time has little impact on the microchannel width. So over long holding time is not needed in hot embossing. Based on the experiment analysis, a series of optimization process parameters is obtained and a fine microfluidic chip is fabricated. The electrophoresis separation experiment are used to verify the microfluidic chip performance after bonding. The results show that 100bp-ladder DNA sample can be separated in less than 5 min successfully.

Identifying EGFR-Expressed Cells and Detecting EGFR Multi-Mutations at Single-Cell Level by Microfluidic Chip

EGFR mutations companion diagnostics have been proved to be crucial for the efficacy of tyrosine kinase inhibitor targeted cancer therapies. To uncover multiple mutations occurred in minority of EGFR-mutated cells,which may be covered by the noises from majority of unmutated cells, is currently becoming an urgent clinical requirement. Here we present the validation of a microfluidic-chip-based method for detecting EGFR multimutations at single-cell level. By trapping and immunofluorescently imaging single cells in specifically designed silicon microwells, the EGFR-expressed cellswere easily identified. By in situ lysing single cells, the cell lysates of EGFR-expressed cells were retrieved without cross-contamination. Benefited from excluding the noise from cells without EGFR expression, the simple and cost-effective Sanger's sequencing, but not the expensive deep sequencing of the whole cell population, was used to discover multi-mutations. We verified the new method with precisely discovering three most important EGFR drugrelated mutations from a sample in which EGFR-mutated cells only account for a small percentage of whole cell population. The microfluidic chip is capable of discovering not only the existence of specific EGFR multi-mutations,but also other valuable single-cell-level information: on which specific cells the mutations occurred, or whether different mutations coexist on the same cells. This microfluidic chip constitutes a promising method to promote simple and cost-effective Sanger's sequencing to be a routine test before performing targeted cancer therapy.

Junction matters in hydraulic circuit bio-design of microfluidics

Microfluidic channels are at micrometer scales;thus,their fluid flows are laminar,resulting in the linear dependence of pressure drop on flow rate in the length of the channel.The ratio of the pressure drop to flow rate,referred to as resistance,depends on channel size and dynamic viscosity.Usually,a microfluidic chip is analogous to an electric circuit in design,but the design is adjusted to optimize channel size.However,whereas voltage loss is negligible at the nodes of an electric circuit,hydraulic pressure drops at the nodes of microfluidic chips by a magnitude are comparable to the pressure drops in the straight channels.Here,we prove by experiment that one must fully consider the pressure drops at nodes so as to accurately design a precise microfluidic chip.In the process,we numerically calculated the pressure drops at hydraulic nodes and list their resistances in the range of flows as concerned.We resorted to machine learning to fit the calculated results for complex junctions.Finally,we obtained a library of node resistances for common junctions and used them to design three established chips that work for single-cell analysis and for precision allocation of solutes(in gradient and averaging concentration microfluidic networks).Endothelial cells were stimulated by generating concentrations of adriamycin hydrochloride from the last two microfluidic networks,and we analyzed the response of endothelial cells.The results indicate that consideration of junction resistances in design calculation brings experimental results closer to the design values than usual.This approach may therefore contribute to providing a platform for the precise design of organ chips.

Wave-shaped microfluidic chip assisted point-of-care testing for accurate and rapid diagnosis of infections

Background:Early diagnosis and classification of infections increase the cure rate while decreasing complications,which is significant for severe infections,especially for war surgery.However,traditional methods rely on laborious operations and bulky devices.On the other hand,point-of-care(POC)methods suffer from limited robustness and accuracy.Therefore,it is of urgent demand to develop POC devices for rapid and accurate diagnosis of infections to fulfill on-site militarized requirements.Methods:We developed a wave-shaped microfluidic chip(WMC)assisted multiplexed detection platform(WMC-MDP).WMC-MDP reduces detection time and improves repeatability through premixing of the samples and reaction of the reagents.We further combined the detection platform with the streptavidin–biotin(SA-B)amplified system to enhance the sensitivity while using chemiluminescence(CL)intensity as signal readout.We realized simultaneous detection of C-reactive protein(CRP),procalcitonin(PCT),and interleukin-6(IL-6)on the detection platform and evaluated the sensitivity,linear range,selectivity,and repeatability.Finally,we finished detecting 15 samples from volunteers and compared the results with commercial ELISA kits.Results:Detection of CRP,PCT,and IL-6 exhibited good linear relationships between CL intensities and concentrations in the range of 1.25–40μg/ml,0.4–12.8 ng/ml,and 50–1600 pg/ml,respectively.The limit of detection of CRP,PCT,and IL-6 were 0.54μg/ml,0.11 ng/ml,and 16.25 pg/ml,respectively.WMC-MDP is capable of good adequate selectivity and repeatability.The whole detection procedure takes only 22 min that meets the requirements of a POC device.Results of 15 samples from volunteers were consistent with the results detected by commercial ELISA kits.Conclusions:WMC-MDP allows simultaneous,rapid,and sensitive detection of CRP,PCT,and IL-6 with satisfactory selectivity and repeatability,requiring minimal manipulation.However,WMC-MDP takes advantage of being a microfluidic device showing the coefficients of variation less than 10%enabling WMC-MDP to be a type of point-of-care testing(POCT).Therefore,WMC-MDP provides a promising alternative to POCT of multiple biomarkers.We believe the practical application of WMC-MDP in militarized fields will revolutionize infection diagnosis for soldiers.

Microfluidic chips for the endothelial biomechanics and mechanobiology of the vascular system

Endothelial cells arranged on the vessel lumen are constantly stimulated by blood flow,blood pressure and pressureinduced cyclic stretch.These stimuli are sensed through mechanical sensory structures and converted into a series of functional responses through mechanotransduction pathways.The process will eventually affect vascular health.Therefore,there has been an urgent need to establish in vitro endothelial biomechanics and mechanobiology of models,which reproduce three-dimensional structure vascular system.In recent years,the rapid development in microfluidic technology makes it possible to replicate the key structural and functionally biomechanical characteristics of vessels.Here,we summarized the progress of microfluidic chips used for the investigation of endothelial biomechanics and mechanobiology of the vascular system.Firstly,we elucidated the contribution of shear stress and circumferential stress,to vascular physiology.Then,we reviewed some applications using microfluidic technology in angiogenesis and vasculogenesis,endothelial permeability and mechanotransduction,as well as the blood-brain barrier under these physical forces.Finally,we discussed the future obstacles in terms of the development and application of microfluidic vascular chips.

A 3D-printed microfluidic gradient concentration chip for rapid antibiotic-susceptibility testing

The rise of antibiotic resistance as one of the most serious global public health threats has necessitated the timely clinical diagnosis and precise treatment of deadly bacterial infections.To identify which types and doses of antibiotics remain effective for fighting against multi-drug-resistant pathogens,the development of rapid and accurate antibiotic-susceptibility testing(AST)is of primary importance.Conventional methods for AST in well-plate formats with disk diffusion or broth dilution are both labor-intensive and operationally tedious.The microfluidic chip provides a versatile tool for evaluating bacterial AST and resistant behaviors.In this paper,we develop an operationally simple,3D-printed microfluidic chip for AST which automatically deploys antibiotic concentration gradients and fluorescence intensity-based reporting to ideally reduce the report time for AST to within 5 h.By harnessing a commercially available,digital light processing(DLP)3D printing method that offers a rapid,high-precision microfluidic chip-manufacturing capability,we design and realize the accurate generation of on-chip antibiotic concentration gradients based on flow resistance and diffusion mechanisms.We further demonstrate the employment of the microfluidic chip for the AST of E.coli to representative clinical antibiotics of three classes:ampicillin,chloramphenicol,and kanamycin.The determined minimum inhibitory concentration values are comparable to those reported by conventional well-plate methods.Our proposed method demonstrates a promising approach for realizing robust,convenient,and automatable AST of clinical bacterial pathogens.

Simultaneous laser-induced fluorescence and contactless-conductivity detection for microfluidic chip

A combined detection system involving simultaneous LIF and contacfless-conductometric measurements at the same place of the microfluidic chip was described. The LIF measurement was designed according to the confocal principle and a moveable contactless-conduetivity detector was used in C^4D. Both measurements were mutually independent and advantageous in analyses of mixtures. Various experimental parameters affecting the response were examined and optimized. The performances were demonstrated by simultaneous detection of Rhodamine B. And the results showed that the combined detection system could be used sensitively and reliably.

MODELING OF LASER MACHINING ON POLYMETHYL METHACRYLATE TO FABRICATE MICROFLUIDIC CHIP

The use of a CO2 laser system for fabrication of microfluidic chip on polymethyl methacrylate （PMMA） is presented to reduce fabrication cost and time of chip. The grooving process of the laser system and a model for the depth of microchannels are investigated. The relations between the depth of laser-cut channels and the laser beam power, velocity or the number of passes of the beam along the same channel are evaluated. In the experiments, the laser beam power varies from 0 to 50 W, the laser beam scanning velocity varies from 0 to 1 000 mm/s and the passes vary in the range of 1 to 10 times. Based on the principle of conservation of energy, the influence of the laser beam velocity, the laser power and the number of groove passes are examine. Considering the grooving interval energy loss, a modified mathematical model has been obtained and experimental data show good agreement with the theoretical model. This approach provides a simple way of predicting groove depths. The system provides a cost alternative of the other methods and it is especially useful on research work of rnicrofluidic prototyping due to the short cycle time of production.

A novel heating area design of temperature-jump microfluidic chip for synchrotron radiation solution X-ray scattering

In this paper, we present a novel design scheme of temperature-jump(T-jump) area for microfluidic device.Numerical simulation and experimental research of thermal characteristics of the solution in microchannels is completed.Numerical simulation of the temperature-jump microchannel is analyzed to study the heat transfer characteristics by comparing performance of three proposed configurations.Calculation of the power requirement is discussed in the dimensional design of microheater. Temperature-sensitive fluorescent dye is applied to investigate the temperature field of microchannel heated by a designed microheater. It is found that the T-jump microfluidic device can provide rapid heating for solutions with strong convection heat transfer ability.

Fabrication of Microfluidic Fiber Chip Detection System

The diameter of the excitation beam was decreased greatly by integrating the fiber on the microfluidic chip as light propagation medium.The coupling efficiency of the fiber was improved with optical fiber collimation device coupling beam. The chip was placed in the darkroom to avoid the interference of the external light.The cost of the instrument was decreased with a high brightness blue LED as excitation source;the performance of the system was valuated by the determination of FITC fluorescein with a minimum detectable concentration of 2.2×10^(-8) mol/L,the Signal-to-Noise Ratio (SNR) S/N=5.The correlation coefficient of the detection system within the range of 1.8×10^(-7) mol/L～4×10^(-5)mol/L was 0.9972.

Impacts of Wettability on Immiscible Fluid Flow Pattern-Microfluidic Chip Experiment

Immiscible fluid flow is frequently found in resource recovery or soil remediation. The flow pattern in the porous media is affected by the wettability of pore surface. In this study, quartz substrates and microfluidic chips are treated by silica nanoparticles(SNP) and triethoxy(octyl)silane(TES) to fabricate the water-wet and oil-wet surfaces. The wettability of the treated-surface is measured in term of contact angle for several combinations of surrounding fluid and a liquid droplet. The effect of the wettability on the fluid flow pattern is explored by injecting oil and water alternately into the surface-treated microfluidic chips. The results reveal that the SNP-coated quartz substrate shows strong water-wet property and the TES coating makes water-repellent/oil-wet surfaces. In addition, it is found that the maximum and minimum oil and water saturation during alternate injection of oil and water depends on the surface wettability of the microfluidic chips. The characteristics of the pore-scale fluid flow pattern are also described.

Integrated Carbon Nanotubes Electrodes in Microfluidic Chip via MWPCVD

An on-chip electrochemical detector for microfluidic chips was described, based on integrated carbon nanotube （CNT） electrodes directly onto the chip substrate through microwave plasma chemical vapor deposition （MWPCVD）. The attractive performance of the integrated CNT electrodes was demonstrated for the amperometric detection of sucrose, glucose and D-fructose. The integrated CNT electrodes showed stronger electrocatalytic activity than gold electrodes.

Microfluidic Chip Method for Multi-SNPs Genotyping in Individual Risk Assessment of Micronutrient Deficiency

Research on nutrigenomics has accumulated sufficient data in the past two decades that have dem on strated phe no types of single n ucleotide polymorphisms (SNPs) betwee n healthy and micronutrient-deficient populations. For instance, Zhang et al. showed that the genes MTHFR C677T, MTRR A66G, and MTR A2756G were the genetic factors resp on sible for low absorptio n and bioavailability of vitamins such as folate, B6/ and B12. It has also been reported that these nutrients are closely associated with the prevale nee of neural tube defects in newborn infants。