Heterogeneous Integration of Acoustic Microextraction with an Optoelectronic Sensor on Glass for Nucleic Acid Testing.

Lab-on-a-chip systems (LOCs), characterized by their high sensitivity, low sample consumption, and portability, have significantly advanced the field of on-site testing. Despite the evolution of integrated LOCs from qualitative to quantitative analyses, on-chip full integration of sample preparation, purification, and multiplexed detection remains a challenge. Here, we propose a strategy for the heterogeneous integration of a set of complementary metal oxide semiconductor-compatible devices including acoustic resonator, thin-film resistors, and temperature/photosensors as a new type of LOC for nucleic acid testing (NAT). Programmed acoustic streaming-based particles and fluid manipulations largely simplify the nucleic acid extraction process including cell lysis, nucleic acid capture, and elution. The design of the acoustic microextraction module and extraction process was thoroughly studied. Benefitted by the microelectromechanical system approach, the conventional mechanical actions and complex flow control are avoided, which enables a compact hand-held NAT instrument without complicated peripherals. Validation experiments conducted on plasma-harboring mutations in the epidermal growth factor receptor (EGFR) gene confirmed the robustness of the system, achieving an impressive nucleic acid (NA) extraction efficiency of approximately 90% within 5 min and a limit of detection of the target NA in the plasma of 1 copy/µL.

Convolutional Neural Network-Driven Impedance Flow Cytometry for Accurate Bacterial Differentiation.

Impedance flow cytometry (IFC) has been demonstrated to be an efficient tool for label-free bacterial investigation to obtain the electrical properties in real time. However, the accurate differentiation of different species of bacteria by IFC technology remains a challenge owing to the insignificant differences in data. Here, we developed a convolutional neural networks (ConvNet) deep learning approach to enhance the accuracy and efficiency of the IFC toward distinguishing various species of bacteria. First, more than 1 million sets of impedance data (comprising 42 characteristic features for each set) of various groups of bacteria were trained by the ConvNet model. To improve the efficiency for data analysis, the Spearman correlation coefficient and the mean decrease accuracy of the random forest algorithm were introduced to eliminate feature interaction and extract the opacity of impedance related to the bacterial wall and membrane structure as the predominant features in bacterial differentiation. Moreover, the 25 optimized features were selected with differentiation accuracies of >96% for three groups of bacteria (bacilli, cocci, and vibrio) and >95% for two species of bacilli (Escherichia coli and Salmonella enteritidis), compared to machine learning algorithms (complex tree, linear discriminant, and K-nearest neighbor algorithms) with a maximum accuracy of 76.4%. Furthermore, bacterial differentiation was achieved on spiked samples of different species with different mixing ratios. The proposed ConvNet deep learning-assisted data analysis method of IFC exhibits advantages in analyzing a huge number of data sets with capacity for extracting predominant features within multicomponent information and will bring about progress and advances in the fields of both biosensing and data analysis.

Acoustofluidic manipulation for submicron to nanoparticles.

Particles, ranging from submicron to nanometer scale, can be broadly categorized into biological and non-biological types. Submicron-to-nanoscale bioparticles include various bacteria, viruses, liposomes, and exosomes. Non-biological particles cover various inorganic, metallic, and carbon-based particles. The effective manipulation of these submicron to nanoparticles, including their separation, sorting, enrichment, assembly, trapping, and transport, is a fundamental requirement for different applications. Acoustofluidics, owing to their distinct advantages, have emerged as a potent tool for nanoparticle manipulation over the past decade. Although recent literature reviews have encapsulated the evolution of acoustofluidic technology, there is a paucity of reports specifically addressing the acoustical manipulation of submicron to nanoparticles. This article endeavors to provide a comprehensive study of this topic, delving into the principles, apparatus, and merits of acoustofluidic manipulation of submicron to nanoparticles, and discussing the state-of-the-art developments in this technology. The discourse commences with an introduction to the fundamental theory of acoustofluidic control and the forces involved in nanoparticle manipulation. Subsequently, the working mechanism of acoustofluidic manipulation of submicron to nanoparticles is dissected into two parts, dominated by the acoustic wave field and the acoustic streaming field. A critical analysis of the advantages and limitations of different acoustofluidic platforms in nanoparticles control is presented. The article concludes with a summary of the challenges acoustofluidics face in the realm of nanoparticle manipulation and analysis, and a forecast of future development prospects.

Enrichment of Aggregation-Induced Emission Aggregates Using Acoustic Streaming Tweezers in Microfluidics for Trace Human Serum Albumin Detection.

Aggregation-dependent brightness (ADB) indirectly limits the in vitro performance of a pure aggregation-induced emission (AIE) probe in many ways; thus, controlling the aggregation state of the AIE probe is helpful for detecting an object of interest. Many studies are focused on the molecule design of the AIE probes, while less efforts have been made for the control of the aggregation of the AIEs. Here, an acoustic streaming tweezer (AST) generated using a gigahertz bulk acoustic wave resonator was applied to manipulate the aggregation status of the AIE probe and further enhance their performance for human serum albumin (HSA) detection. As the trapping size of the AST matches the working size of the AIE probe, the streaming can enrich and accumulate AIE nanoparticles, which then further trigger larger aggregates. Due to the ADB effect, the fluorescence intensity strongly increased, and thus, the detection limit of HSA was reduced to 0.5 µg/mL, which is low enough for kidney disease detection. Such an AST-assisted ADB strategy is potentially applicable to other AIE probes and can work as a portable choice for the biomedical detection.

On-Chip Arbitrary Manipulation of Single Particles by Acoustic Resonator Array.

Effective and arbitrary manipulation of particles in liquid has attracted substantial interest. Acoustic tweezers, a new and promising tool, exhibit high biocompatibility, universality, and precision but lack arbitrariness. In this work, we report a gigahertz (GHz) bulk acoustic streaming tweezer (AST)-based micro-manipulation platform capable of efficiently translating acoustic energy to fluid kinetic energy, creating a controllable, quick-response, and stable flow field and precisely, arbitrarily, and universally manipulating a single particle to move like a microrobot. Through controlling the radio frequency signals applied on these resonators, the intensity and direction of the acoustic streaming flow can be quickly and arbitrarily adjusted. Consequently, the particle dispersed at the bottom can be arbitrarily and steadily driven along the predesigned route to the target position by the acoustic streaming drag force (ASF). We utilized four resonators cooperated as a work group to manipulate single SiO2 particles to complete nearly uniform linear motions and U-shaped motions, as well as playing billiards and exploring a maze, demonstrating the enormous potential of this GHz AST-based single-particle manipulation platform for separation, assembly, sensing, enriching, transporting, and so forth.

Hypersound-Assisted Size Sorting of Microparticles on Inkjet-Patterned Protein Films.

Hydrodynamic approaches are important for biomedical diagnostics, chemical analysis, and a broad range of industrial applications. Size-based separation and sorting is an important tool for these applications. We report the integration of hypersound technology with patterned protein films to provide efficient sorting of microparticles based on particle charge and size. We employed a hypersonic resonator for the acoustic streaming of the fluidic system to generate microvortices that exert drag forces on the objects on the surface that are dictated by their radius of curvature. We demonstrate a size-based sorting of anionic silica particles using protein patterns and gradients fabricated using attractive cationic and repulsive anionic proteins.

Mechanism and stability investigation of a nozzle-free droplet-on-demand acoustic ejector.

This paper investigates the mechanism of a new acoustic micro-ejector using a Lamb wave transducer array, which can stably generate picoliter (pL) droplet jetting without nozzles. With eight transducers arranged as an octagon array, droplets are ejected based on the mechanism of combined acoustic pressure waves and acoustic streaming. The acoustic focusing area is designed as a line at the liquid center, which is the key factor for a large working range of liquid height. The experimental results show that the ejector can produce uniform water droplets of 22 µm diameter (5.6 pL in volume) continuously at a rate of 0.33 kHz with high ejection stability, owing to a large liquid height window and high acoustic wave frequency. By delivering precise ∼pL droplets without clogging issues, the acoustic ejector has great potential for demanding biochemical applications.

Resistive pulse sensing device with embedded nanochannel (nanochannel-RPS) for label-free biomolecule and bionanoparticle analysis.

This paper reports an IC-compatible method for fabricating a PDMS-based resistive pulse sensing (RPS) device with embedded nanochannel (nanochannel-RPS) for label-free analysis of biomolecules and bionanoparticles, such as plasmid DNAs and exosomes. Here, a multilayer lithography process was proposed to fabricate the PDMS mold for the microfluidic device, comprising a bridging nanochannel, as the sensing gate. RPS was performed by placing the sensing and excitation electrodes symmetrically upstream and downstream of the sensing gate. In order to reduce the noise level, a reference electrode was designed and placed beside the excitation electrode. To demonstrate the feasibility of the proposed nanochannel-RPS device and sensing system, polystyrene micro- and nanoparticles with diameters of 1µm and 300 nm were tested by the proposed device with signal-to-noise ratios (SNR) ranging from 9.1-30.5 and 2.2-5.9, respectively. Furthermore, a nanochannel with height of 300 nm was applied for 4 kb plasmid DNA detection, implying the potential of the proposed method for label-free quantification of nanoscale biomolecules. Moreover, HeLa cell exosomes, known as a well-studied subtype of extracellular vesicles, were measured and analyzed by their size distribution. The result of the resistive pulse amplitude corresponded well to that of nanoparticle tracking analysis (NTA). The proposed nanochannel-RPS device and the sensing strategy are not only capable of label-free analysis for nanoscale biomolecules and bionanoparticles, but are also cost-effective for large-scale manufacturing.

In-Line Detection with Microfluidic Bulk Acoustic Wave Resonator Gas Sensor for Gas Chromatography.

A microfluidic film bulk acoustic wave resonator gas sensor (mFBAR) adapted specifically as an in-line detector in gas chromatography was described. This miniaturized vapor sensor was a non-destructive detector with very low dead volume (0.02 µL). It was prepared by enclosing the resonator in a microfluidic channel on a chip with dimensions of only 15 mm × 15 mm × 1 mm. The device with polymer coating showed satisfactory performance in the detection of organophosphorus compound, demonstrating a very low detection limit (a dozen parts per billion) with relatively short response time (about fifteen seconds) toward the simulant of chemical warfare agent, dimethyl methylphosphonate. The in-line detection of the mFBAR sensor with FID was constructed and employed to directly measure the concentration profile on the solid surface by the mFBAR with the controlled concentration profile in the mobile phase at the same time. The difference of peak-maximum position between mobile phase and solid phase could be a convenient indicator to measure mass transfer rate. With the response of the mFBAR and FID obtained in one injection, an injection mass-independent parameter can be calculated and used to identify the analyte of interest.

Label-Free and Simultaneous Mechanical and Electrical Characterization of Single Plant Cells Using Microfluidic Impedance Flow Cytometry.

Despite that single-cell-type-level analyses have been extensively conducted on animal models to gain new insights into complex biological processes; the unique biological and physiological properties of plant cells have not been widely studied at single-cell resolution. In this work, an electrical impedance flow cytometry was fabricated based on microfluidics with constriction microchannel to simultaneously characterize the mechanical and electrical properties of single plant cells. Protoplasts from two model plant species, the herbaceous Arabidopsis thaliana and the woody Populus trichocarpa, could be readily discriminated by their respective mechanical traits, but not by electrical impedance. On the contrary, overexpression of a red fluorescent protein on plasma membrane resulted in changes in cell electrical impedance instead of cell deformability. During primary cell wall (PCW) regeneration, this extracellular layer outside of protoplasts introduced dramatic variations in both mechanical and electrical properties of single plant cells. Furthermore, the effects of auxin, an essential phytohormone regulating PCW reformation, were validated on this platform. Taken together, our results revealed a novel application of microfluidic impedance flow cytometry in the field of plant science to simultaneously characterize dual biophysical properties at single-cell resolution, which could be further developed as a powerful and reliable tool for plant cell phenotyping and cell fate specification.

A Highly Aligned Nanowire-Based Strain Sensor for Ultrasensitive Monitoring of Subtle Human Motion.

Achieving highly accurate responses to external stimuli during human motion is a considerable challenge for wearable devices. The present study leverages the intrinsically high surface-to-volume ratio as well as the mechanical robustness of nanostructures for obtaining highly-sensitive detection of motion. To do so, highly-aligned nanowires covering a large area were prepared by capillarity-based mechanism. The nanowires exhibit a strain sensor with excellent gauge factor (≈35.8), capable of high responses to various subtle external stimuli (≤200 µm deformation). The wearable strain sensor exhibits also a rapid response rate (≈230 ms), mechanical stability (1000 cycles) and reproducibility, low hysteresis (<8.1%), and low power consumption (<35 µW). Moreover, it achieves a gauge factor almost five times that of microwire-based sensors. The nanowire-based strain sensor can be used to monitor and discriminate subtle movements of fingers, wrist, and throat swallowing accurately, enabling such movements to be integrated further into a miniaturized analyzer to create a wearable motion monitoring system for mobile healthcare.

Rapid response flexible humidity sensor for respiration monitoring using nano-confined strategy.

Development of wearable devices for continuous respiration monitoring is of great importance for evaluating human health. Here, we propose a new strategy to achieve rapid respiration response by confining conductive polymers into 1D nanowires which facilitates the water molecules absorption/desorption and maximizes the sensor response to moisture. The nanowires arrays were fabricated through a low-cost nanoscale printing approach on flexible substrate. The nanoscale humidity sensor shows a high sensitivity (5.46%) and ultrafast response (0.63 s) when changing humidity between 0% and 13% and can tolerate 1000 repetitions of bending to a curvature radius of 10 mm without influencing its performance. Benefited by its fast response and low power assumption, the humidity sensor was demonstrated to monitor human respiration in real time. Different respiration patterns including normal, fast and deep respiration can be distinguished accurately.

Miniature Gigahertz Acoustic Resonator and On-Chip Electrochemical Sensor: An Emerging Combination for Electroanalytical Microsystems.

Performance of electroanalytical lab-on-a-chip devices is often limited by the mass transfer of electroactive species toward the electrode surface, due to the difficulty in applying external convection. This article describes the powerful signal enhancement attained with a 2.54 GHz miniature acoustic resonator integrated with an electrochemical device in a miniaturized cell. Acoustic resonator and an on-chip gold thin-film three-electrode electrochemical cell were arranged facing each other inside a structured poly(methyl methacrylate) chamber. Cyclic voltammetric and chronoamperometric responses of 1 mM ferrocene-methanol were recorded under resonator's actuation at powers ranging from 0 to 1 W. Finite element analysis was carried out to study the sono-electroanalytical process. Acoustic resonator's actuation greatly enhances the mass transport of electroactive species toward the electrode surface. The diffusion limited cyclic voltammetric and chronoamperometric currents increase around 10 and 20 times, respectively, with an input power of 1 W compared to those recorded under stagnant conditions. The improvement in electroanalytical process is mainly associated with acoustic resonator's vibration induced fluid streaming. The advantages of a miniaturized acoustic resonator, including the submillimeter small size, amenability for mass fabrication, cost effectiveness, low energy consumption, as well as outstanding enhancement of coupled electrochemical processes, will enable the production of highly sensitive compact electroanalytical devices.

Hierarchical assembly of gold nanorod stripe patterns for sensing and cells alignment.

Hierarchical assemblies of nanomaterial superstructures with controlled orientation affords a multitude of novel properties of plasmonics and broad applications. Yet constructing multi-functional superstructures with nanoparticles positioned in desired locations remains challenging. Herein, gold nanorods (GNRs) assembled in stripe patterns with controlled orientation and structures in millimeter scale for versatile application have been achieved. Applications of patterned GNRs in sensing enhancement and engineering mammalian cells alignment are investigated experimentally. The performance of patterned GNRs in surface enhanced Raman scattering (SERS) and electrical sensing are found in orientational dependence. The SERS signals of vertically arranged GNR arrays exhibit double the folder intensity than those horizontally arranged. In contrast, the horizontally arranged GNRs exhibit twice as much electrical conductivity. The system is further explored to pattern mammalian cells. For the first time, we reveal the nanostructured topography of GNR confined cells to a specific region, and direct the adhesion and extension of living cells, which opens up broad applications in tissue engineering and biosensing.

Programmable multi-DNA release from multilayered polyelectrolytes using gigahertz nano-electromechanical resonator.

BACKGROUND: Controllable and multiple DNA release is critical in modern gene-based therapies. Current approaches require complex assistant molecules for combined release. To overcome the restrictions on the materials and environment, a novel and versatile DNA release method using a nano-electromechanical (NEMS) hypersonic resonator of gigahertz (GHz) frequency is developed. RESULTS: The micro-vortexes excited by ultra-high frequency acoustic wave can generate tunable shear stress at solid-liquid interface, thereby disrupting molecular interactions in immobilized multilayered polyelectrolyte thin films and releasing embedded DNA strands in a controlled fashion. Both finite element model analysis and experiment results verify the feasibility of this method. The release rate and released amount are confirmed to be well tuned. Owing to the different forces generated at different depth of the films, release of two types of DNA molecules with different velocities is achieved, which further explores its application in combined gene therapy. CONCLUSIONS: Our research confirmed that this novel platform based on a nano-electromechanical hypersonic resonator works well for controllable single and multi-DNA release. In addition, the unique features of this resonator such as miniaturization and batch manufacturing open its possibility to be developed into a high-throughput, implantable and site targeting DNA release and delivery system.

Controlled and Tunable Loading and Release of Vesicles by Using Gigahertz Acoustics.

Controllable exchange of molecules between the interior and the external environment of vesicles is critical in drug delivery and micro/nano-reactors. While many approaches exist to trigger release from vesicles, controlled loading remains a challenge. Herein, we show that gigahertz acoustic streaming generated by a nanoelectromechanical resonator can control the loading and release of cargo into and from vesicles. Polymer-shelled vesicles showed loading and release of molecules both in solution and on a solid substrate. We observed deformation of individual giant unilamellar vesicles and propose that the shear stress generated by gigahertz acoustic streaming induces the formation of transient nanopores, with diameters on the order of 100 nm, in the vesicle membranes. This provides a non-invasive method to control material exchange across membranes of different types of vesicles, which could allow site-specific release of therapeutics and controlled loading into cells, as well as tunable microreactors.

Dual-Mode Gas Sensor Composed of a Silicon Nanoribbon Field Effect Transistor and a Bulk Acoustic Wave Resonator: A Case Study in Freons.

In this paper, we develop a novel dual-mode gas sensor system which comprises a silicon nanoribbon field effect transistor (Si-NR FET) and a film bulk acoustic resonator (FBAR). We investigate their sensing characteristics using polar and nonpolar organic compounds, and demonstrate that polarity has a significant effect on the response of the Si-NR FET sensor, and only a minor effect on the FBAR sensor. In this dual-mode system, qualitative discrimination can be achieved by analyzing polarity with the Si-NR FET and quantitative concentration information can be obtained using a polymer-coated FBAR with a detection limit at the ppm level. The complementary performance of the sensing elements provides higher analytical efficiency. Additionally, a dual mixture of two types of freons (CFC-113 and HCFC-141b) is further analyzed with the dual-mode gas sensor. Owing to the small size and complementary metal-oxide semiconductor (CMOS)-compatibility of the system, the dual-mode gas sensor shows potential as a portable integrated sensing system for the analysis of gas mixtures in the future.

Hypersonic Poration: A New Versatile Cell Poration Method to Enhance Cellular Uptake Using a Piezoelectric Nano-Electromechanical Device.

Efficient delivery of genes and therapeutic agents to the interior of the cell is critical for modern biotechnology. Herein, a new type of chemical-free cell poration method-hypersonic poration-is developed to improve the cellular uptake, especially the nucleus uptake. The hypersound (≈GHz) is generated by a designed piezoelectric nano-electromechanical resonator, which directly induces normal/shear stress and "molecular bombardment" effects on the bilayer membranes, and creates reversible temporal nanopores improving the membrane permeability. Both theory analysis and cellular uptake experiments of exogenous compounds prove the high delivery efficiency of hypersonic poration. Since target molecules in cells are accumulated with the treatment, the delivered amount can be controlled by tuning the treatment time. Furthermore, owing to the intrinsic miniature of the resonator, localized drug delivery at a confined spatial location and tunable arrays of the resonators that are compatible with multiwell plate can be achieved. The hypersonic poration method shows great delivery efficacy combined with advantage of scalability, tunable throughput, and simplification in operation and provides a potentially powerful strategy in the field of molecule delivery, cell transfection, and gene therapy.

Conductive polymer nanowire gas sensor fabricated by nanoscale soft lithography.

Resistive devices composed of one-dimensional nanostructures are promising candidates for the next generation of gas sensors. However, the large-scale fabrication of nanowires is still challenging, which restricts the commercialization of such devices. Here, we report a highly efficient and facile approach to fabricating poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS) nanowire chemiresistive gas sensors by nanoscale soft lithography. Well-defined sub-100 nm nanowires are fabricated on silicon substrate, which facilitates device integration. The nanowire chemiresistive gas sensor is demonstrated for NH3 and NO2 detection at room temperature and shows a limit of detection at ppb level, which is compatible with nanoscale PEDOT:PSS gas sensors fabricated with the conventional lithography technique. In comparison with PEDOT:PSS thin-film gas sensors, the nanowire gas sensor exhibits higher sensitivity and a much faster response to gas molecules.

Novel Gas Sensor Arrays Based on High-Q SAM-Modified Piezotransduced Single-Crystal Silicon Bulk Acoustic Resonators.

This paper demonstrates a novel micro-size (120 µm × 200 µm) piezoelectric gas sensor based on a piezotransduced single-crystal silicon bulk acoustic resonator (PSBAR). The PSBARs operate at 102 MHz and possess high Q values (about 2000), ensuring the stability of the measurement. A corresponding gas sensor array is fabricated by integrating three different self-assembled monolayers (SAMs) modified PSBARs. The limit of detection (LOD) for ethanol vapor is demonstrated to be as low as 25 ppm with a sensitivity of about 1.5 Hz/ppm. Two sets of identification code bars based on the sensitivities and the adsorption energy constants are utilized to successfully discriminate isopropanol (IPA), ethanol, hexane and heptane vapors at low and high gas partial pressures, respectively. The proposed sensor array shows the potential to form a portable electronic nose system for volatile organic compound (VOC) differentiation.