Advances in microfluidic-based DNA methylation analysis

DNA methylation has been extensively investigated in recent years,not least because of its known relationship with various diseases.Progress in analytical methods can greatly increase the relevance of DNA methylation studies to both clinical medicine and scientific research.Microflu-idic chips are excellent carriers for molecular analysis,and their use can provide improvements from multiple aspects.On-chip molecular analysis has received extensive attention owing to its advantages of portability,high throughput,low cost,and high efficiency.In recent years,the use of novel microfluidic chips for DNA methylation analysis has been widely reported and has shown obvious superiority to conventional methods.In this review,wefirst focus on DNA methylation and its applications.Then,we discuss advanced microfluidic-based methods for DNA methylation analysis and describe the great progress that has been made in recent years.Finally,we summarize the advantages that microfluidic technology brings to DNA methylation analysis and describe several challenges and perspectives for on-chip DNA methylation analysis.This review should help researchers improve their understanding and make progress in developing microfluidic-based methods for DNA methylation analysis.

Particle distributions in Lamb wave based acoustofluidics

Acoustic streaming enabled by a Lamb wave resonator(LWR)is efficient for particle trapping and enrichment in microfluidic channels.However,because Lamb waves combine the features of bulk acoustic waves and surface acoustic waves,the resulting acoustic streaming in the LWR occurs in multiple planes,and the particle flow behavior in this acoustofluidic system is largely unknown.Reported here are numerical simulations and laboratory experiments conducted to investigate the boundary conditions for particle motion inside a microvortex induced by an LWR.Upon dynamic capture,the particles’trajectories become orbital paths within an acoustic vortex.The suspended particles encounter two distinct acoustic phenomena,i.e.,the drag force resulting from acoustic streaming and the acoustic radiation force,which exert forces in various directions on the particles.When the acoustic radiation force and the fluid drag force are dominant for large and small particles in a mixed solution,respectively,the large particles reside within the vortex while the small particles remain at its periphery.Conversely,when the acoustic radiation force is dominant for both types of particles,the distribution pattern is reversed.

Nanostructure enabled extracellular vesicles separation and detection

Extracellular vesicles(EVs)have recently attracted significant research attention owing to their important biological functions,including cell-to-cell communication.EVs are a type of membrane vesicles that are secreted into the extracellular space by most types of cells.Several biological biomolecules found in EVs,such as proteins,microRNA,and DNA,are closely related to the pathogenesis of human malignancies,making EVs valuable biomarkers for disease diagnosis,treatment,and prognosis.Therefore,EV separation and detection are prerequisites for providing important information for clinical research.Conventional separation methods suffer from low levels of purity,as well as the need for cumbersome and prolonged operations.Moreover,detection methods require trained operators and present challenges such as high operational expenses and low sensitivity and specificity.In the past decade,platforms for EV separation and detection based on nanostructures have emerged.This article reviews recent advances in nanostructure-based EV separation and detection techniques.First,nanostructures based on membranes,nanowires,nanoscale deterministic lateral displacement,and surface modification are presented.Second,high-throughput separation of EVs based on nanostructures combined with acoustic and electric fields is described.Third,techniques combining nanostructures with immunofluorescence,surface plasmon resonance,surface-enhanced Raman scattering,electrochemical detection,or piezoelectric sensors for high-precision EV analysis are summarized.Finally,the potential of nanostructures to detect individual EVs is explored,with the aim of providing insights into the further development of nanostructure-based EV separation and detection techniques.

Tunable microfluidic chip for single-cell deformation study

Microfluidic phenotyping methods have been of vital importance for cellular characterization,especially for evaluating single cells.In order to study the deformability of a single cell,we devised and tested a tunable microfluidic chip-based method.A pneumatic polymer polydimethylsiloxane(PDMS)membrane was designed and fabricated abutting a single-cell trapping structure,so the cell could be squeezed controllably in a lateral direction.Cell contour changes under increasing pressure were recorded,enabling the deformation degree of different types of single cell to be analyzed and compared using computer vision.This provides a new perspective for studying mechanical properties of cells at the single cell level.

State-of-the-art and recent developments in micro/nanoscale pressure sensors for smart wearable devices and health monitoring systems

Small-sized,low-cost,and high-sensitivity sensors are required for pressure-sensing applications because of their critical role in consumer electronics,automotive applications,and industrial environments.Thus,micro/nanoscale pressure sensors based on micro/nanofabrication and micro/nanoelectromechanical system technologies have emerged as a promising class of pressure sensors on account of their remarkable miniaturization and performance.These sensors have recently been developed to feature multifunctionality and applicability to novel scenarios,such as smart wearable devices and health monitoring systems.In this review,we summarize the major sensing principles used in micro/nanoscale pressure sensors and discuss recent progress in the development of four major categories of these sensors,namely,novel material-based,flexible,implantable,and selfpowered pressure sensors.

Recent advances in micro/nanoscale intracellular delivery

Intracellular delivery enables the efficient drug delivery into various types of cells and has been a long-term studied topics in modern biotechnology.Targeted delivery with improved delivery efficacy requires considerable requirements.This process is a critical step in many cellular-level studies,such as cellular drug therapy,gene editing delivery,and a series of biomedical research applications.The emergence of micro-and nanotechnology has enabled themore accurate and dedicated intracellular delivery,and it is expected to be the next generation of controlled delivery with unprecedented flexibility.This review focuses on several represented micro-and nanoscale physical approaches for cell membrane disruption-based intracellular delivery and discusses the mechanisms,advantages,and challenges of each approach.We believe that the deeper understanding of intracellular delivery at such lowdimensionwould help the research community to develop more powerful delivery technologies for biomedical applications.

Theoretical and experimental characterizations of gigahertz acoustic streaming in microscale fluids

Even as gigahertz(GHz) acoustic streaming has developed into a multi-functional platform technology for biochemical applications, including ultrafast microfluidic mixing, microparticle operations, and cellar or vesicle surgery, its theoretical principles have yet to be established. This is because few studies have been conducted on the use of such high frequency acoustics in microscale fluids. Another difficulty is the lack of velocimetry methods for microscale and nanoscale fluidic streaming. In this work, we focus on the basic aspects of GHz acoustic streaming,including its micro-vortex generation principles, theoretical model, and experimental characterization technologies. We present details of a weak-coupled finite simulation that represents our current understanding of the GHz-acoustic-streaming phenomenon. Both our simulation and experimental results show that the GHzacoustic-induced interfacial body force plays a determinative role in vortex generation. We carefully studied changes in the formation of GHz acoustic streaming at different acoustic powers and flow rates. In particular,we developed a microfluidic-particle-image velocimetry method that enables the quantification of streaming at the microscale and even nanoscale. This work provides a full map of GHz acoustofluidics and highlights the way to further theoretical study of this topic.

Manipulations of micro/nanoparticles using gigahertz acoustic streaming tweezers

Contactless acoustic manipulation of micro/nanoscale particles has attracted considerable attention owing to its near independence of the physical and chemical properties of the targets,making it universally applicable to almost all biological systems.Thin-film bulk acoustic wave(BAW)resonators operating at gigahertz(GHz)frequencies have been demonstrated to generate localized high-speed microvortices through acoustic streaming effects.Benefitting from the strong drag forces of the high-speed vortices,BAW-enabled GHz acoustic streaming tweezers(AST)have been applied to the trapping and enrichment of particles ranging in size from micrometers to less than 100 nm.However,the behavior of particles in such 3D microvortex systems is still largely unknown.In this work,the particle behavior(trapping,enrichment,and separation)in GHz AST is studied by theoretical analyses,3D simulations,and microparticle tracking experiments.It is found that the particle motion in the vortices is determined mainly by the balance between the acoustic streaming drag force and the acoustic radiation force.This work can provide basic design principles for AST-based lab-on-a-chip systems for a variety of applications.

A combined virtual impactor and field-effect transistor microsystem for particulate matter separation and detection

Ambient suspended particulate matter(PM)(primarily with particle diameter 2.5m or less,i.e.,PM2.5)can adversely affect ecosystems and human health.Currently,optical particle sensors based on light scattering dominate the portable PM sensing market.However,the light scattering method has poor adaptability to different-sized PM and adverse environmental conditions.Here,we design and develop a portable PM sensing microsystem that consists of a micromachined virtual impactor(VI)for particle separation,a thermophoretic deposition chip for particle collection,and an extended-gate field-effect transistor(FET)for particle analysis.This system can realize on-site separation,collection,and analysis of aerosol particles without being influenced by environmental factors.In this study,the design of the VI is thoroughly analyzed by numerical simulation,and mixtures of different-sized silicon dioxide(SiO2)particles are used in an experimental verification of the performance of the VI and FET.Considering the low cost and compact design of the whole system,the proposed PM analysis microsystem has potential for PM detection under a wide range of conditions,such as heavily polluted industrial environments and for point-of-need outdoor and indoor air quality monitoring.

PEDOT:PSS:From conductive polymers to sensors

PEDOT:PSS conductive polymers have received tremendous attention over the last two decades owing to their high conductivity,ease of processing,and biocompatibility.As a flexible versatile material,PEDOT:PSS can be developed into various forms and has had a significant impact on emerging sensing applications.This review covers the development of PEDOT:PSS from material to physical sensors.We focus on the morphology of PEDOT:PSS in the forms of aqueous dispersions,solid films,and hydrogels.Manufacturing processes are summarized,including coating,printing,and lithography,and there is particular emphasis on nanoimprinting lithography that enables the production of PEDOT:PSS nanowires with superior sensing performance.Applications to various physical sensors,for humidity,temperature,pressure,and strain,are demonstrated.Finally,we discuss the challenges and propose new directions for the development of PEDOT:PSS.

A prototype portable instrument employing micro-preconcentrator and FBAR sensor for the detection of chemical warfare agents

The presence of chemical warfare agents(CWAs)in the environment is a serious threat to human safety,but there are many problems with the currently available detection methods for CWAs.For example,gas chromatography–mass spectrometry cannot be used for in-field detection owing to the rather large size of the equipment required,while commercial sensors have the disadvantages of low sensitivity and poor selectivity.Here,we develop a portable gas sensing instrument for CWA detection that consists of a MEMSfabricated micro-preconcentrator(μPC)and a film bulk acoustic resonator(FBAR)gas sensor.The μPC is coated with a nanoporous metal–organic framework material to enrich the target,while the FBAR provides rapid detection without the need for extra carrier gas.Dimethyl methylphosphonate(DMMP),a simulant of the chemical warfare agent sarin,is used to test the performance of the instrument.Experimental results show that the μPC provides effective sample pretreatment,while the FBAR gas sensor has good sensitivity to DMMP vapor.The combination of μPC and FBAR in one instrument gives full play to their respective advantages,reducing the limit of detection of the analyte.Moreover,both the μPC and the FBAR are fabricated using a CMOS-compatible approach,and the prototype instrument is compact in size with high portability and thus has potential for application to in-field detection of CWAs.

Investigation of sorptive interactions between volatile organic compounds and supramolecules at dynamic oscillation using bulk acoustic wave resonator virtual sensor arrays

Supramolecules are considered as promising materials for volatile organic compounds(VOCs)sensing applications.The proper understanding of the sorption process taking place in host-guest interactions is critical in improving the pattern recognition of supramolecules-based sensing arrays.Here,we report a novel approach to investigate the dynamic host-guest recognition process by employing a bulk acoustic wave(BAW)resonator capable of producing multiple oscillation amplitudes and simultaneously recording multiple responses to VOCs.Self-assembled monolayers(SAMs)ofβ-cyclodextrin(β-CD)were modified on four BAW sensors to demonstrate the gas-surface interactions regarding oscillation amplitude and SAM length.Based on the method,a virtual sensor array(VSA)type electronic nose(e-nose)can be realized by pattern recognition of multiple responses at different oscillation amplitudes of a single sensor.VOCs analysis was realized respectively by using principal component analysis(PCA)for individual VOC identification and linear discriminant analysis(LDA)for VOCs mixtures classification.

Recent advances in micro detectors for micro gas chromatography

Micro gas chromatography(μGC) has been continuously gaining attention since the last century owing to multiple favorable characteristics, such as its small size, low power consumption and minimal production and maintenance costs.μGC has the potential to provide practical solutions to emerging analytical challenges in security, health,and environment. In this review, we summarize recent advances in micro detectors for μGC, including the study of the miniaturization of conventional detectors and the development of novel detectors for μGC chromatography.

Manipulation of single cells via a Stereo Acoustic Streaming Tunnel(SteAST)

At the single-cell level,cellular parameters,gene expression and cellular function are assayed on an individual but not population-average basis.Essential to observing and analyzing the heterogeneity and behavior of these cells/clusters is the ability to prepare and manipulate individuals.Here,we demonstrate a versatile microsystem,a stereo acoustic streaming tunnel,which is triggered by ultrahigh-frequency bulk acoustic waves and highly confined by a microchannel.We thoroughly analyze the generation and features of stereo acoustic streaming to develop a virtual tunnel for observation,pretreatment and analysis of cells for different single-cell applications.3D reconstruction,dissociation of clusters,selective trapping/release,in situ analysis and pairing of single cells with barcode gel beads were demonstrated.To further verify the reliability and robustness of this technology in complex biosamples,the separation of circulating tumor cells from undiluted blood based on properties of both physics and immunity was achieved.With the rich selection of handling modes,the platform has the potential to be a full-process microsystem,from pretreatment to analysis,and used in numerous fields,such as in vitro diagnosis,high-throughput single-cell sequencing and drug development.