A titanium dioxide nanorod array as a high-affinity nano-bio interface of a microfluidic device for efficient capture of circulating tumor cells

Nanomaterials show promising opportunities to address clinical problems （such as insufficient capture of circulating tumor cells; CTCs） via the high surface area-to-volume ratio and high affinity for biological cells. However, how to apply these nanomaterials as a nano-bio interface in a microfluidic device for efficient CTC capture with high specificity remains a challenge. In the present work, we first found that a titanium dioxide （TiO2） nanorod array that can be conveniently prepared on multiple kinds of substrates has high affinity for tumor cells. Then, the TiO2 nanorod array was vertically grown on the surface of a microchannel with hexagonally patterned Si micropillars via a hydrothermal reaction, forming a new kind of a micro-nano 3D hierarchically structured microfluidic device. The vertically grown TiO2 nanorod array was used as a sensitive nano-bio interface of this 3D hierarchically structured microfluidic device, which showed high efficiency of CTC capture （76.7% ± 7.1%） in an artificial whole-blood sample.

Highly precise Ti/Pt/Cr/Au thin-film temperature sensor embedded in a microfluidic device

A multilayer(Ti/Pt/Cr/Au)resistive temperature sensor was proposed and investigated to precisely measure the temperature characteristic in microfluidic devices.The Ti/Pt/Cr/Au sensor was fabricated by direct current(DC)sputtering,vacuum evaporation and liftoff process.The thermal annealing test was conducted in the temperature range of 200-800℃for obtaining an appropriate property of the multilayer.Based on the experimental results,400℃was selected as the experimental annealing temperature for the Ti/Pt/Cr/Au layer.The redistribution of structural imperfections and recrystallization promote the density and adhesion of multilayer during the annealing process.With the annealing temperature rising,the annealing process leads to through-thickness migration of chromium and partial depletion of the adhesive layer.The Ti also diffuses into the Pt,which makes the interface disappear.Nevertheless,the layer remains continuous.The temperature coefficient of resistance(TCR)of the sensors was investigated through the microfluidic testing system.The excellent stability and sensitivity of the Ti/Pt/Cr/Au thin-film temperature sensor are verified.Furthermore,the capability of the Ti/Pt/Cr/Au thin-film temperature sensor detecting the sudden temperature change caused by bubble effect is very meaningful to the microfluidic devices.

A microfluidic device for accurate detection of hs-cTnI

This device is aimed at ensuring that the sample is uniformly and equivalently reacted with the antibody on the NC membrane in each test when the microfluidic liquid system is introduced to the chip.In this study,the developed microfluidic chip can avoid the presence of the sample and conjugate pads in the chip,while the precision of the chro matography system can be greatly improved using the same particles,NC membrane and antibody alongside the traditional strip.The results,taking the detection of cTnI as an example,revealed that the coefficient of variation(CV)is controlled within 4%,while the maximum record of the contrast chromatographic reagent strip can reach 15%.Additionally,the detection sensitivity can maintain the same order of magnitudes with that of the traditional chromatographic strip.With the results,the determination correlation of the developed microfluidic chip has been greatly improved.In addition,the CV of the chip in this study is greatly improved in comparison with that of the traditional strip.The biggest improvement lies in the mixing between the sample and the microspheres,indicating that this is a new approach to improve the CV of the traditional strip.

Microfluidic Device： A Miniaturized Platform for Chemical Reactions

Microfluidic devices, as a new miniaturized platform stemming from the field of micro-electromechanical sys-tems, have been used in many disciplines. In the field of chemical reactions, microfluidic device-based microreac-tors have shown great promise in building new chemical technologies and processes with increased speed and reli- ability and reduced sample consumption and cost. This technology has also become a new and effective tool for precise, high-throughput, and automatic analysis of chemical synthesis processes. Compared with conventional chemical laboratory batch methodologies, microfluidic reactors have a number of features, such as high mixing ef- ficiency, short reaction time, high heat-transfer coefficient, small reactant volume, controllable residence time, and high surface-to-volume ratio, among others. Combined with recent advances in microfluidic devices for chemical reactions, this review aims to give an overview of the features and applications of microfluidic devices in the field of chemical synthesis. It also aims to stimulate the development of microfluidic device applications in the field of chemical reactions.

Hierarchical Porous Polymer Beads Prepared by Polymerization-induced Phase Separation and Emulsion-template in a Microfluidic Device

Porous polymer beads（PPBs） containing hierarchical bimodal pore structure with gigapores and meso-macropores were prepared by polymerization-induced phase separation（PIPS） and emulsion-template technique in a glass capillary microfluidic device（GCMD）. Fabrication procedure involved the preparation of water-in-oil emulsion by emulsifying aqueous solution into the monomer solution that contains porogen. The emulsion was added into the GCMD to fabricate the（water-in-oil）-in-water double emulsion droplets. The flow rate of the carrier continuous phase strongly influenced the formation mechanism and size of droplets. Formation mechanism transformed from dripping to jetting and size of droplets decreased from 550 μm to 250 μm with the increase in flow rate of the carrier continuous phase. The prepared droplets were initiated for polymerization by on-line UV-irradiation to form PPBs. The meso-macropores in these beads were generated by PIPS because of the presence of porogen and gigapores obtained from the emulsion-template. The pore morphology and pore size distribution of the PPBs were investigated extensively by scanning electron microscopy and mercury intrusion porosimetry（MIP）. New pore morphology was formed at the edge of the beads different from traditional theory because of different osmolarities between the water phase of the emulsion and the carrier continuous phase. The morphology and proportion of bimodal pore structure can be tuned by changing the kind and amount of porogen.

Utilizing a microfluidic device to enrich and fluorescently detect circulating tumor cells

Circulating tumor cells(CTCs)are cancer cells that have propagated from primary tumor sites,spreading into the bloodstream as the cellular origin of fatal metastasis,and to secondary tumor sites.Capturing and analyzing CTCs is a kind of‘‘liquid biopsy'of the tumor that provides information about cancer changes over time and tailoring treatment[1].CTC enrichment and detection remains technologically challenging due to their extremely low concentra-

Microfluidic devices as model platforms of CNS injury-ischemia to study axonal regeneration by regulating mitochondrial transport and bioenergetic metabolism

Central nervous system(CNS)neurons typically fail to regenerate their axons after injury leading to neurological impairment.Axonal regeneration is a highly energy-demanding cellular program that requires local mitochondria to supply most energy within injured axons.Recent emerging lines of evidence have started to reveal that injury-triggered acute mitochondrial damage and local energy crisis contribute to the intrinsic energetic restriction that accounts for axon regeneration failure in the CNS.Characterizing and reprogramming bioenergetic signaling and mitochondrial maintenance after axon injury-ischemia is fundamental for developing therapeutic strategies that can restore local energy metabolism and thus facilitate axon regeneration.Therefore,establishing reliable and reproduc-ible neuronal model platforms is critical for assessing axonal energetic metabolism and regeneration capacity after injury-ischemia.In this focused methodology article,we discuss recent advances in applying cutting-edge microflu-idic chamber devices in combination with state-of-the-art live-neuron imaging tools to monitor axonal regeneration,mitochondrial transport,bioenergetic metabolism,and local protein synthesis in response to injury-ischemic stress in mature CNS neurons.

In vitro engineered models of neurodegenerative diseases

Neurodegeneration is a catastrophic process that develops progressive damage leading to functional andstructural loss of the cells of the nervous system and is among the biggest unavoidable problems of our age.Animalmodels do not reflect the pathophysiology observed in humans due to distinct differences between the neuralpathways,gene expression patterns,neuronal plasticity,and other disease-related mechanisms in animals andhumans.Classical in vitro cell culture models are also not sufficient for pre-clinical drug testing in reflecting thecomplex pathophysiology of neurodegenerative diseases.Today,modern,engineered techniques are applied to developmulticellular,intricate in vitro models and to create the closest microenvironment simulating biological,biochemical,and mechanical characteristics of the in vivo degenerating tissue.In THIS review,the capabilities and shortcomings ofscaffold-based and scaffold-free techniques,organoids,and microfluidic models that best reflect neurodegeneration invitro in the biomimetic framework are discussed.

Microfluidics-based assay on the effects of microenvironmental geometry and aqueous flow on bacterial adhesion behaviors

Abstract A new microfluidic system with four different microchambers （a circle and three equilateral concave polygons） was designed and fabricated using poly（dimethylsiloxane） （PDMS） and the soft lithography method. Using this microfluidic device at six flow rates （5, 10, 20, 30, 40, and 50 μL/h）, the effects of microenvironmental geometry and aqueous flow on bacterial adhesion behaviors were investigated. Escherichia coli HB101 pGLO, which could produce a green fluorescent protein induced by L-arabinose, was utilized as the model bacteria. The results demonstrated that bacterial adhesion was significantly related to culture time, microenvironment geometry, and aqueous flow rates. Adhered bacterial density increased with the culture time. Initially, the adhesion occurred at the microchamber sides, and then the entire chamber was gradually covered with increased culture time. Adhesion densities in the side zones were larger than those in the center zones because of the lower shearing force in the side zone. Also, the adhesion densities in the complex chambers were larger than those in the simple chambers. At low flow rates, the orientation of adhered bacteria was random and disorderly. At high flow rates, bacterial orientation became close to the streamline and oriented toward the flow direction; All these results implied that bacterial adhesion tended to occur in complicated aqueous flow areas.The present study provided an on-chip flow system for physiological behavior of biological cells, as well as provided a strategic cue for the prevention of bacterial infection and biofilm formation.

Current overview of induced pluripotent stem cell-based blood-brain barrier-on-a-chip

BACKGROUND Induced pluripotent stem cells(iPSCs)show great ability to differentiate into any tissue,making them attractive candidates for pathophysiological investigations.The rise of organ-on-a-chip technology in the past century has introduced a novel way to make in vitro cell cultures that more closely resemble their in vivo environments,both structural and functionally.The literature still lacks consensus on the best conditions to mimic the blood-brain barrier(BBB)for drug screening and other personalized therapies.The development of models based on BBB-on-achip using iPSCs is promising and is a potential alternative to the use of animals in research.AIM To analyze the literature for BBB models on-a-chip involving iPSCs,describe the microdevices,the BBB in vitro construction,and applications.METHODS We searched for original articles indexed in PubMed and Scopus that used iPSCs to mimic the BBB and its microenvironment in microfluidic devices.Thirty articles were identified,wherein only 14 articles were finally selected according to the inclusion and exclusion criteria.Data compiled from the selected articles were organized into four topics:(1)Microfluidic devices design and fabrication;(2)characteristics of the iPSCs used in the BBB model and their differentiation conditions;(3)BBB-on-a-chip reconstruction process;and(4)applications of BBB microfluidic three-dimensional models using iPSCs.RESULTS This study showed that BBB models with iPSCs in microdevices are quite novel in scientific research.Important technological advances in this area regarding the use of commercial BBB-on-a-chip were identified in the most recent articles by different research groups.Conventional polydimethylsiloxane was the most used material to fabricate in-house chips(57%),whereas few studies(14.3%)adopted polymethylmethacrylate.Half the models were constructed using a porous membrane made of diverse materials to separate the channels.iPSC sources were divergent among the studies,but the main line used was IMR90-C4 from human fetal lung fibroblast(41.2%).The cells were differentiated through diverse and complex processes either to endothelial or neural cells,wherein only one study promoted differentiation inside the chip.The construction process of the BBB-on-a-chip involved previous coating mostly with fibronectin/collagen Ⅳ(39.3%),followed by cell seeding in single cultures(36%)or co-cultures(64%)under controlled conditions,aimed at developing an in vitro BBB that mimics the human BBB for future applications.CONCLUSION This review evidenced technological advances in the construction of BBB models using iPSCs.Nonetheless,a definitive BBB-on-a-chip has not yet been achieved,hindering the applicability of the models.

Axonal regeneration and sprouting as a potential therapeutic target for nervous system disorders

Nervous system disorders are prevalent health issues that will only continue to increase in frequency as the population ages.Dying-back axonopathy is a hallmark of many neurologic diseases and leads to axonal disconnection from their targets,which in turn leads to functional impairment.During the course of many of neurologic diseases,axons can regenerate or sprout in an attempt to reconnect with the target and restore synapse function.In amyotrophic lateral sclerosis(ALS),distal motor axons retract from neuromuscular junctions early in the disease-course before significant motor neuron death.There is evidence of compensatory motor axon sprouting and reinnervation of neuromuscular junctions in ALS that is usually quickly overtaken by the disease course.Potential drugs that enhance compensatory sprouting and encourage reinnervation may slow symptom progression and retain muscle function for a longer period of time in ALS and in other diseases that exhibit dying-back axonopathy.There remain many outstanding questions as to the impact of distinct disease-causing mutations on axonal outgrowth and regeneration,especially in regards to motor neurons derived from patient induced pluripotent stem cells.Compartmentalized microfluidic chambers are powerful tools for studying the distal axons of human induced pluripotent stem cells-derived motor neurons,and have recently been used to demonstrate striking regeneration defects in human motor neurons harboring ALS disease-causing mutations.Modeling the human neuromuscular circuit with human induced pluripotent stem cells-derived motor neurons will be critical for developing drugs that enhance axonal regeneration,sprouting,and reinnervation of neuromuscular junctions.In this review we will discuss compensatory axonal sprouting as a potential therapeutic target for ALS,and the use of compartmentalized microfluidic devices to find drugs that enhance regeneration and axonal sprouting of motor axons.

3D tumor model biofabrication

Animal models have been extensively used in cancer pathology studies and drug discovery.These models,however,fail to reflect the complex human tumor microenvironment and do not allow for high-throughput drug screening in more human-like physiological conditions.Three-dimensional(3D)cancer models present an alternative to automated high-throughput cancer drug discovery and oncology.In this review,we highlight recent technology innovations in building 3D tumor models that simulate the complex human tumor microenvironment and responses of patients to treatment.We discussed various biofabrication technologies,including 3D bioprinting techniques developed for characterizing tumor progression,metastasis,and response to treatment.

Theoretical, Simulation and Experimental Analysis of Microfluidic Droplet Generation and Recovering Process with Applications in Frying Oil Assessments

The research on microfluidic droplet size prediction has been extensive and fruitful, while the droplet deforming process has been seldom studied. In this paper, a frying-oil-assessing microfluidic device was designed to study the droplet deforming and recovering processes, which were dominated by channel geometry, flow rates,sheath flow viscosity and interfacial tension of the two phases. Theoretical expressions of the deforming process and its extreme value were obtained for the first time, supported by simulation and experiments. Theoretical,simulation and experimental results indicated that the steady-state droplet length could be a useful parameter for frying oil assessment.

Distance-Based Detection of Ag^(+) with Gold Nanoparticles-Coated Microfluidic Paper

In this study,we developed a microfluidic paper analysis device(μPAD)for distance-based detection of Ag^(+)in water.TheμPAD was manufactured by wax printing method on filter paper.Then,a layer of gold nanoparticles(AuNPs)was deposited and ascorbic acid was printed on the channel.In the detection,Ag^(+)was reduced by ascorbic acid and coated on the surface of the AuNPs on the channel,forming Au@Ag core/shell nanoparticles.Based on the capillary flow principle,diff erent concentrations of Ag^(+)formed diff erent distances of color ribbons.Thus,quantitative detection of Ag^(+)can be achieved by measuring the distance of the color ribbon.The detection limit of this method was as low as 1 mg·L^(-1)within 15 min and the interference of common metal ions in water can be eliminated.In conclusion,this method had successfully realized the leap from colorimetry to direct reading,realizing fast read and easy manipulation with low-cost.

Low-temperature optothermal nanotweezers

Optical tweezers that rely on laser irradiation to capture and manipulate nanoparticles have provided powerful tools for biological and biochemistry studies.However,the existence of optical diffraction-limit and the thermal damage caused by high laser power hinder the wider application of optical tweezers in the biological field.For the past decade,the emergence of optothermal tweezers has solved the above problems to a certain extent,while the auxiliary agents used in optothermal tweezers still limit their biocompatibility.Here,we report a kind of nanotweezers based on the sign transformation of the thermophoresis coefficient of colloidal particles in low-temperature environment.Using a self-made microfluidic refrigerator to reduce the ambient temperature to around 0℃in the microfluidic cell,we can control a single nanoparticle at lower laser power without adding additional agent solute in the solution.This novel optical tweezering scheme has provided a new path for the manipulation of inorganic nanoparticles as well as biological particles.

Fabrication of Polymersomes with Controllable Morphologies through Dewetting w/o/w Double Emulsion Droplets

In this work, monodisperse giant polymersomes are fabricated by dewetting of water-in-oil-in-water double emulsion droplets which are assembled by amphiphilic block copolymer molecules in a microfluidic device. The dewetting process can be tuned by solvation between solvent and amphiphilic block copolymer to get polymersomes with controllable morphology. Good solvent （chloroform and toluene） hinders dewetting process of double emulsion droplets and gets acornlike polymersomes or patched polymersomes. On the other hand, poor solvent （hexane） accelerates the dewetting process and achieves complete separation of inner water phase from oil phase to form complete bilayer polymersomes. In addition, twin polymersomes with bilayer membrane structure are formed by this facile method. The formation mechanism for different polymersomes is discussed in detail.

Functional poly(carboxybetaine methacrylate)coated paper sensor for high efficient and multiple detection of nutrients in fruit

Rapid and simultaneous in situ detection of multi-components is extremely crucial for the real-time monitoring of nutrients in fruits.Herein,a facile and user-friendly poly(carboxybetaine methacrylate)-coated paper-based microfluidic device(pCBMA-μPAD)has been exploited to synchronously identify and semi-quantify vitamin C.glucose,sucro se and fructose in fruits.The pCBMA was successfully grafted from the surface of paper sensor using a co nvenient and robust method,which was confirmed by Fourier transform infrared spectroscopy(FT-IR)and X-ray photoelectron spectrometry(XPS).The superior hydrophilicity and ultra-low fouling of pCBMA endowed the pCBMA-μPAD with remarkably rapid response(3 min),high sensitivity,good linear relationship and low detection limit(LOD)(vitamin C:y=33.809+5.175 x,R^2=0.993,LOD=0.179 mmol/L;glucose:y=-0.113+30.0661 g(x),R^2=0.988,LOD=0.095 mmol/L;sucrose:y--5.334+34.858 lg(x),R^2=0.996,LOD=0.097 mmol/L;fructose:y=4.996+23.325 lg(x),R^2=0.994,LOD=0.140 mmol/L).Furthermore,satisfactory results were yielded in the detection of these nutrients in 9 fruits,which were much agreed well with those obtained by spectrophotometry.Such a portable and versatile pCBMA-μPAD will pro foundly shape the future of food analysis,especially for the assessment of food quality and nutrition in the process of agricultural production and marketing.

An openable artificial intestinal tract system for the in vitro evaluation of medicines

In vitro drug screening systems for pharmacological targets have been studied as substitutes for whole-animal experiments.Cultured cells or tissues provide promising substitution models when coupled with technological innovations in micro total analysis systems.In this study,we focus on an intestinal drug absorption assay,as the oral route is most frequently used for drug administration.Pharmacological studies have reported the development of artificial vessels that include tubular structures.However,it is difficult to observe the insides of these tubes in situ.To address this problem,we developed a micro-device that uses a pneumatic balloon actuator(PBA)to open and close an artificial intestinal tract.A human colon carcinoma cell line(Caco-2)was cultivated on the flat surface of the micro-device for 7 days to form the inner cellular layer of an artificial intestinal tract with which to evaluate drug transport.The artificial intestinal tract was completely actuated from a flat plate to a circular tube via a PBA with a pressure of 65 kPa,and drugs were perfused at a flow rate of 0.05 mL min^(−1) into the tubular artificial intestinal tract for 1 h.Using the openable artificial intestinal tract,the in vitro absorption of calcein and Texas Red were successfully estimated as models of hydrophilic and hydrophobic drugs,respectively.The artificial intestinal tract enables the effective evaluation of the in vitro intestinal absorption of drug candidates and contributes to the reduction of costs incurred during the initial stage of drug development.