Microfluidic systems for axonal growth and regeneration research

Damage to the adult mammalian central nervous system （CNS） often results in persistent neurological deficits with limited recovery of functions. The past decade has seen in- creasing research efforts in neural regeneration research with the ultimate goal of achieving functional recovery. Many studies have focused on prevention of further neural damage and restoration of functional connections that are com- promised after iniurY or pathological damage.

Engineering organoid microfluidic system for biomedical and health engineering:A review

In recent years,organoid technology,i.e.,in vitro three-dimensional(3D)tissue culture,has attracted increasing attention in biomedical engineering.Organoids are cell complexes induced by differentiation of stem cells or organ-progenitor cells in vitro using 3D culture technology.They can replicate the key structural and functional characteristics of the target organs in vivo.With the opening up of this new field of health engineering,there is a need for engineering-system approaches to the production,control,and quantitative analysis of organoids and their microenvironment.Traditional organoid technology has limitations,including lack of physical and chemical microenvironment control,high heterogeneity,complex manual operation,imperfect nutritional supply system,and lack of feasible online analytical technology for the organoids.The introduction of microfluidic chip technology into organoids has overcome many of these limitations and greatly expanded the scope of applications.Engineering organoid microfluidic system has become an interdisciplinary field in biomedical and health engineering.In this review,we summarize the development and culture system of organoids,discuss how microfluidic technology has been used to solve the main technical challenges in organoid research and development,and point out new opportunities and prospects for applications of organoid microfluidic system in drug development and screening,food safety,precision medicine,and other biomedical and health engineering fields.

Microfluidic System for Synthesis of Trigonal Selenium Nanowires

A microfluidic system was developed for the synthesis of trigonal selenium（t-Se） nanowires, which was composed of a glass microchip coupled with a poly（methyl methacrylate）（PMMA） microchip. In the glass microchip, amorphous selenium（a-Se） colloid was prepared by reducing selenious acid with an excess amount of hydrazine at a temperature of 100 ℃. In the coupled PMMA microchip, a-Se was transformed into more stable t-Se seeds via sonication at room temperature. The residence time of the reactants in both microchips was optimized by varying the dimension and length of the microchannel each. The t-Se nanowires were formed by anisotropic growth of selenium crystallite during sonication and aging under the assistance of β-cyclodextrin（β-CD）. Various stages of the nanowires＇ growth were investigated. The as-synthesized products were characterized by powder X-Ray diffraction（XRD）, Raman spectroscopy, scanning electron microscopy（SEM）, transmission electron microscopy（TEM） and selected-area electron diffraction（SAED）.

A Microfluidic System with Active Mixing for Improved Real-Time Isothermal Amplification

To improve the performance of real-time recombinase polymerase amplification(RPA),a microfluidic system with active mixing is developed to optimize the reaction dynamics.Instead of adopting a single typical reaction chamber,a specific reactor including a relatively large chamber in center with two adjacent zig-zag channels at two sides is integrated into the microfluidic chip.Active mixing is achieved by driving the viscous reagent between the chamber and the channel back and forth periodically with an outside compact peristaltic pump.To avoid reagent evapora-tion,one end of the reactor is sealed with paraffin oil.A hand-held companion device is developed to facilitate real-time RPA amplification within 20 min.The whole area of the reactor is heated with a resistance heater to provide uniform reaction temperature.To achieve real-time monitoring,a compact fluorescence detection module is integrated into the hand-held device.A smartphone with custom application software is adopted to control the hand-held device and display the real-time fluorescence curves.The performances of two cases with and without active on-chip mixing are compared between each other by detecting African swine fever viruses.It has been demonstrated that,with active on-chip mixing,the amplification efficiency and detection sensitivity can be signifi-cantly improved.

Femtosecond laser-induced nanoparticle implantation into flexible substrate for sensitive and reusable microfluidics SERS detection

Surface-enhanced Raman spectroscopy(SERS)microfluidic system,which enables rapid detection of chemical and biological analytes,offers an effective platform to monitor various food contaminants and disease diagnoses.The efficacy of SERS microfluidic systems is greatly dependent on the sensitivity and reusability of SERS detection substrates to ensure repeated use for prolonged periods.This study proposed a novel process of femtosecond laser nanoparticle array(NPA)implantation to achieve homogeneous forward transfer of gold NPA on a flexible polymer film and accurately integrated it within microfluidic chips for SERS detection.The implanted Au-NPA strips show a remarkable electromagnetic field enhancement with the factor of 9×108 during SERS detection of malachite green(MG)solution,achieving a detection limit lower than 10 ppt,far better than most laser-prepared SERS substrates.Furthermore,Au-NPA strips show excellent reusability after several physical and chemical cleaning,because of the robust embedment of laser-implanted NPA in flexible substrates.To demonstrate the performance of Au-NPA,a SERS microfluidic system is built to monitor the online oxidation reaction between MG/NaClO reactants,which helps infer the reaction path.The proposed method of nanoparticle implantation is more effective than the direct laser structuring technique.It provides better performance for SERS detection,robustness of detection,and substrate flexibility and has a wider range of applications for microfluidic systems without any negative impact.

A crosstalk-free dual-mode sweat sensing system for naked-eye sweat loss quantification via changes in structural reflectance

Sweat loss monitoring is important for understanding the body’s thermoregulation and hydration status,as well as for comprehensive sweat analysis.Despite recent advances,developing a low-cost,scalable,and universal method for the fabrication of colorimetric microfluidics designed for sweat loss monitoring remains challenging.In this study,we propose a novel laserengraved surface roughening strategy for various flexible substrates.This process permits the construction of microchannels that show distinct structural reflectance changes before and after sweat filling.By leveraging these unique optical properties,we have developed a fully laser-engraved microfluidic device for the quantification of naked-eye sweat loss.This sweat loss sensor is capable of a volume resolution of 0.5µL and a total volume capacity of 11µL,and can be customized to meet different performance requirements.Moreover,we report the development of a crosstalk-free dual-mode sweat microfluidic system that integrates an Ag/AgCl chloride sensor and a matching wireless measurement flexible printed circuit board.This integrated system enables the real-time monitoring of colorimetric sweat loss signals and potential ion concentration signals without crosstalk.Finally,we demonstrate the potential practical use of this microfluidic sweat loss sensor and its integrated system for sports medicine via on-body studies.

Microfluidic-oriented synthesis of enriched iridium nanodots/carbon architecture for robust electrocatalytic nitrogen fixation

Electrocatalytic nitrogen reduction reaction(NRR)is considered as a promising candidate to achieve ammonia synthesis because of clean electric energy,moderate reaction condition,safe operating process and harmless by-products.However,the chemical inertness of nitrogen and poor activated capacity on catalyst surface usually produce low ammonia yield and faradic efficiency.Herein,the microfluidic technology is proposed to efficiently fabricate enriched iridium nanodots/carbon architecture.Owing to in-situ co-precipitation reaction and microfluidic manipulation,the iridium nanodots/carbon nanomaterials possess small average size,uniform dispersion,high conductivity and abundant active sites,producing good proton activation and rapid electrons transmission and moderate adsorption/desorption capacity.As a result,the as-prepared iridium nanodots/carbon nanomaterials realize large ammonia yield of 28.73 μg h^(-1) cm^(-2) and faradic efficiency of 9.14%in KOH solution.Moreover,the high ammonia yield of 11.21 μg h^(-1) cm^(-2) and faradic efficiency of 24.30%are also achieved in H_(2)SO_(4) solution.The microfluidic method provides a reference for large-scale fabrication of nano-sized catalyst materials,which may accelerate the progress of electrocatalytic NRR in industrialization field.

3D Microfluidic System for Evaluating Inhibitory Effect of Chinese Herbal Medicine Oldenlandia diffusa on Human Malignant Glioma Invasion Combined with Network Pharmacology Analysis

Objective:To investigate the anti-invasion efficacy of the ethanol extract of Oldenlandia diffusa Will.(EEOD) on a three-dimensional(3D) human malignant glioma(MG) cell invasion and perfusion model based on microfluidic chip culture and the possible mechanism of action of Oldenlandia diffusa Will.(OD).Methods:The comprehensive pharmacodynamic analysis method in this study was based on microfluidic chip 3D cell perfusion culture technology,and the action mechanism of Chinese medicine(CM) on human MG cells was investigated through network pharmacology analysis.First,the components of EEOD were analyzed by ultraperformance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS).Then,cell viability and apoptosis were assessed to determine the optimum concentration of EEOD for invasion experiments,and two-dimensional(2D) migration and invasion abilities of U87 and U251 MG cells were evaluated using scratch wound and Transwell assays.The possible mechanism underlying the effects of EEOD on glioma was analyzed through a network pharmacology approach.Results:Thirty-five compounds of EEOD were detected by UPLC-Q-TOF/MS.EEOD suppressed the viability of MG cells,promoted their apoptosis,and inhibited their migratory and invasive potentials(all P<0.05).Network pharmacology analysis showed that OD inhibited the invasion of MG cells by directly regulating MAPK and Wnt pathways through MAPK,EGFR,MYC,GSK3B,and other targets.The anti-invasion effect of OD was also found to be related to the indirect regulation of microtubule cytoskeleton organization.Conclusion:EEOD could inhibit the invasion of human MG cells,and the anti-invasion mechanism of OD might be regulating MAPK and Wnt signaling pathways and microtubule cytoskeleton organization.

A digital microfluidic single-cell manipulation systemoptimized by extending-depth-of-field device

Microfuidic systems have been widely utilized in high-throughput biology analysis,but thedificulties in iquid manipulation and cell cultivation limit its application.This work has developed a new digital microfluidic(DMF)system for on-demand droplet control.By adopting anextending-depth-of-field(EDoF)phase modulator to the optical system,the entire depth of themicrofluidic channel can be covered in one image without any refocusing process,ensuring that 95%of the particles in the droplet are captured within three shots together with shaking pro-cesses.With this system,suspension droplets are generated and droplets containing only oneyeast cll can be recognized,then each single cell is cultured in the array of the chip.Byobservingtheir growth in cell numbers and the green fluorescence protein(GFP)production via fluorescence imaging,the single cell with the highest production can be identified.The results haveproved the heterogeneity of yeast cells,and showed that the combined system can be applied forrapid single-cell sorting,cultivation,and analysis.

High-throughput microfluidic production of carbon capture microcapsules:fundamentals,applications,and perspectives

In the last three decades,carbon dioxide(CO_(2)) emissions have shown a significant increase from various sources.To address this pressing issue,the importance of reducing CO_(2) emissions has grown,leading to increased attention toward carbon capture,utilization,and storage strategies.Among these strategies,monodisperse microcapsules,produced by using droplet microfluidics,have emerged as promising tools for carbon capture,offering a potential solution to mitigate CO_(2) emissions.However,the limited yield of microcapsules due to the inherent low flow rate in droplet microfluidics remains a challenge.In this comprehensive review,the high-throughput production of carbon capture microcapsules using droplet microfluidics is focused on.Specifically,the detailed insights into microfluidic chip fabrication technologies,the microfluidic generation of emulsion droplets,along with the associated hydrodynamic considerations,and the generation of carbon capture microcapsules through droplet microfluidics are provided.This review highlights the substantial potential of droplet microfluidics as a promising technique for large-scale carbon capture microcapsule production,which could play a significant role in achieving carbon neutralization and emission reduction goals.

Microfluidic thermotaxic selection of highlymotile sperm and in vitro fertilization

The selection of the most motile and functionally competent sperm is an essential basis for in vitro fertilization(IVF)and normal embryonic development.Widely adopted clinical approaches for sperm sample processing intensely rely on centrifugation and wash steps that may induce mechanical damage and oxidative stress to sperm.Although a few microfluidic sperm sorting devices may avoid these adverse effects by exploiting intrinsic guidance mechanisms of sperm swimming,none of these approaches have been fully validated by clinical-grade assessment criteria.In this study,a microfluidic sperm sorting device that enables the selection of highly motile and functional sperm via their intrinsic thermotaxis is presented.Bioinspired by the temperature microenvironment in the fallopian tube during natural sperm selection,a microfluidic device with controllable temperature gradients along the sperm separation channel was designed and fabricated.This study investigated the optimal temperature conditions for human sperm selection and fully characterized thermotaxis-selected sperm with 45 human sperm samples.Results indicated that a temperature range of 35–36.5℃along the separation channel significantly improves human sperm motility rate((85.25±6.28)%vs.(60.72±1.37)%;P=0.0484),increases normal sperm morphology rate((16.42±1.43)%vs.(12.55±0.88)%;P<0.0001),and reduces DNA fragmentation((7.44±0.79)%vs.(10.36±0.72)%;P=0.0485)compared to the nonthermotaxis group.Sperm thermotaxis is species-specific,and selected mouse sperm displayed the highest motility in response to a temperature range of 36–37.5℃ along the separation channel.Furthermore,IVF experiments indicated that the selected sperm permitted an increased fertilization rate and improved embryonic development from zygote to blastocyst.This microfluidic thermotaxic selection approach will be translated into clinical practice to improve the IVF success rate for patients with oligozoospermia and asthenozoospermia.

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.

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.

Microfluidic preparation of surfactant-free ultrafine DAAF with tunable particle size for insensitive initiator explosives

High purity and ultrafine DAAF(u-DAAF)is an emerging insensitive charge in initiators.Although there are many ways to obtain u-DAAF,developing a preparation method with stable operation,accurate control,good quality consistency,equipment miniaturization,and minimum manpower is an inevitable requirement to adapt to the current social technology development trend.Here reported is the microfluidic preparation of u-DAAF with tunable particle size by a passive swirling microreactor.Under the guidance of recrystallization growth kinetics and mixing behavior of fluids in the swirling microreactor,the key parameters(liquid flow rate,explosive concentration and crystallization temperature)were screened and optimized through screening experiments.Under the condition that no surfactant is added and only experimental parameters are controlled,the particle size of recrystallized DAAF can be adjusted from 98 nm to 785 nm,and the corresponding specific surface area is 8.45 m^(2)·g^(-1)to 1.33 m^(2)·g^(-1).In addition,the preparation method has good batch stability,high yield(90.8%-92.6%)and high purity(99.0%-99.4%),indicating a high practical application potential.Electric explosion derived flyer initiation tests demonstrate that the u-DAAF shows an initiation sensitivity much lower than that of the raw DAAF,and comparable to that of the refined DAAF by conventional spraying crystallization method.This study provides an efficient method to fabricate u-DAAF with narrow particle size distribution and high reproducibility as well as a theoretical reference for fabrication of other ultrafine explosives.

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.

Micro-nano-fabrication of green functional materials by multiphase microfluidics for environmental and energy applications

Multiphase microfluidic has emerged as a powerful platform to produce novel materials with tailor-designed functionalities,as microfluidic fabrication provides precise controls over the size,component,and structure of resultant materials.Recently,functional materials with well-defined micro-/nanostructures fabricated by microfluidics find important applications as environmental and energy materials.This review first illustrated in detail how different structures or shapes of droplet and jet templates are formed by typical configurations of microfluidic channel networks and multiphase flow systems.Subsequently,recent progresses on several representative energy and environmental applications,such as water purification,water collecting and energy storage,were overviewed.Finally,it is envisioned that integrating microfluidics and other novel materials will play increasing important role in contributing environmental remediation and energy storage in near future.

Functional microfluidics:theory,microfabrication,and applications

Microfluidic devices are composed of microchannels with a diameter ranging from ten to a few hundred micrometers.Thus,quite a small(10-9–10-18l)amount of liquid can be manipulated by such a precise system.In the past three decades,significant progress in materials science,microfabrication,and various applications has boosted the development of promising functional microfluidic devices.In this review,the recent progress on novel microfluidic devices with various functions and applications is presented.First,the theory and numerical methods for studying the performance of microfluidic devices are briefly introduced.Then,materials and fabrication methods of functional microfluidic devices are summarized.Next,the recent significant advances in applications of microfluidic devices are highlighted,including heat sinks,clean water production,chemical reactions,sensors,biomedicine,capillaric circuits,wearable electronic devices,and microrobotics.Finally,perspectives on the challenges and future developments of functional microfluidic devices are presented.This review aims to inspire researchers from various fields engineering,materials,chemistry,mathematics,physics,and more—to collaborate and drive forward the development and applications of functional microfluidic devices,specifically for achieving carbon neutrality.

Microphysiological systems for modeling gut-organ interaction

The gut is a digestive organ that absorbs nutrients but also plays a vital role in immune response and defense against external compounds.The complex interaction between the gut microbiota and other organs including the immune system of the host has been known in various contexts,yielding the notion of‘axes’between the gut and other organs.While the presence of various gut-organ axes has been reported,the lack of adequate in vitro model systems for studying this interaction has restricted a deeper insight into these phenomena.Recently developed microphysiological systems(MPS),also known as organ-on-a-chip,allow researchers to study complex interactions between diverse organs,and here we provide a review of how recently developed gut-on-a-chip systems are used for building models of various diseases that were difficult to study.

A novel method for fast and continuous preparation of superfine titanium dioxide nanoparticles in microfluidic system

Previously we had developed a microfluidic system that can be easily fabricated by bending a stainless-steel tube into large circular loops.In this study,a fast and continuous preparation method for superfine TiO_(2) nanoparticles(TiO_(2)-NPs)was developed for the aforementioned microfluidic system.The proposed method can yield anatase TiO_(2) in 3.5 min,in contrast to the traditional hydrothermal reaction method,which requires hours or even days.Different reaction conditions,such as reaction temperature(120-200℃),urea concentration(20-100 g/L),and tube length(5-20 m)were investigated.X-ray diffraction and Brunauer-Emmett-Teller analysis indicate that the as-prepared TiO_(2)-NPs have crystalline sizes of 4.1-5.8 nm and specific surface areas of 250.7-330.7 m^(2)/g.Transmission electron microscopy images show that these TiO_(2)-NPs have an even diameter of approximately 5 nm.Moreover,because of their small crystalline sizes and large specific surface areas,most of these as-prepared TiO_(2)-NPs exhibit considerably better absorption and photocatalytic performance with methylene blue than commercial P5 TiO_(2) does.

A fully automated centrifugal microfluidic system for sample-to-answer viral nucleic acid testing

The outbreak of virus-induced infectious diseases poses a global public-health challenge.Nucleic acid amplification testing(NAAT)enables early detection of pandemic viruses and plays a vital role in preventing onward transmission.However,the requirement of skilled operators,expensive instrumentation,and biosafety laboratories has hindered the use of NAAT for screening and diagnosis of suspected patients.Here we report development of a fully automated centrifugal microfluidic system with sample-in-answer-out capability for sensitive,specific,and rapid viral nucleic acid testing.The release of nucleic acids and the subsequent reverse transcription loop-mediated isothermal amplification(RT-LAMP)were integrated into the reaction units of a microfluidic disc.The whole processing steps such as injection of reagents,fluid actuation by rotation,heating and temperature control,and detection of fluorescence signals were carried out automatically by a customized instrument.We validate the centrifugal microfluidic system using oropharyngeal swab samples spiked with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)armored RNA particles.The estimated limit of detection for armored RNA particles is 2 copies per reaction,the throughput is 21 reactions per disc,and the assay sample-to-answer time is approximately 70 min.This enclosed and automated microfluidic system efficiently avoids viral contamination of aerosol,and can be readily adapted for virus detection outside the diagnostic laboratory.