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.

Concentration of Microparticles/Cells Based on an Ultra-Fast Centrifuge Virtual Tunnel Driven by a Novel Lamb Wave Resonator Array.

The µTAS/LOC, a highly integrated microsystem, consolidates multiple bioanalytical functions within a single chip, enhancing efficiency and precision in bioanalysis and biomedical operations. Microfluidic centrifugation, a key component of LOC devices, enables rapid capture and enrichment of tiny objects in samples, improving sensitivity and accuracy of detection and diagnosis. However, microfluidic systems face challenges due to viscosity dominance and difficulty in vortex formation. Acoustic-based centrifugation, particularly those using surface acoustic waves (SAWs), have shown promise in applications such as particle concentration, separation, and droplet mixing. However, challenges include accurate droplet placement, energy loss from off-axis positioning, and limited energy transfer from low-frequency SAW resonators, restricting centrifugal speed and sample volume. In this work, we introduce a novel ring array composed of eight Lamb wave resonators (LWRs), forming an Ultra-Fast Centrifuge Tunnel (UFCT) in a microfluidic system. The UFCT eliminates secondary vortices, concentrating energy in the main vortex and maximizing acoustic-to-streaming energy conversion. It enables ultra-fast centrifugation with a larger liquid capacity (50 µL), reduced power usage (50 mW) that is one order of magnitude smaller than existing devices, and greater linear speed (62 mm/s), surpassing the limitations of prior methods. We demonstrate successful high-fold enrichment of 2 µm and 10 µm particles and explore the UFCT's potential in tissue engineering by encapsulating cells in a hydrogel-based micro-organ with a ring structure, which is of great significance for building more complex manipulation platforms for particles and cells in a bio-compatible and contactless manner.

Low-Voltage High-Frequency Lamb-Wave-Driven Micromotors.

By leveraging the benefits of a high energy density, miniaturization and integration, acoustic-wave-driven micromotors have recently emerged as powerful tools for microfluidic actuation. In this study, a Lamb-wave-driven micromotor is proposed for the first time. This motor consists of a ring-shaped Lamb wave actuator array with a rotor and a fluid coupling layer in between. On a driving mechanism level, high-frequency Lamb waves of 380 MHz generate strong acoustic streaming effects over an extremely short distance; on a mechanical design level, each Lamb wave actuator incorporates a reflector on one side of the actuator, while an acoustic opening is incorporated on the other side to limit wave energy leakage; and on electrical design level, the electrodes placed on the two sides of the film enhance the capacitance in the vertical direction, which facilitates impedance matching within a smaller area. As a result, the Lamb-wave-driven solution features a much lower driving voltage and a smaller size compared with conventional surface acoustic-wave-driven solutions. For an improved motor performance, actuator array configurations, rotor sizes, and liquid coupling layer thicknesses are examined via simulations and experiments. The results show the micromotor with a rotor with a diameter of 5 mm can achieve a maximum angular velocity of 250 rpm with an input voltage of 6 V. The proposed micromotor is a new prototype for acoustic-wave-driven actuators and demonstrates potential for lab-on-a-chip applications.

Microfluidic confined acoustic streaming vortex for liposome synthesis.

Liposomes have garnered significant attention owing to their favorable characteristics as promising carriers. Microfluidic based hydrodynamic flow focusing, or micro-mixing approaches enable precise control of liposome size during their synthesis due to the comparable size scale. However, current microfluidic approaches still have issues such as high flow rate dependency, complex chip structures, and ease of clogging. Herein, we present a novel microfluidic platform for size-tunable liposome synthesis based on an ultra-high-frequency acoustic resonator. By designing the shape and orientation of the acoustic resonator in the three-phase laminar flow, it combined the features of both hydrodynamic flow focusing and rapid micro-mixing. The distribution of lipid precursor solution in laminar flow and the mixing conditions could be regulated by the confined acoustic streaming vortex. We successfully synthesize liposomes with adjustable sizes and narrow size distributions. Notably, this platform regulates the product size by adjusting only the input power, which is less dependent on the flow rate. Furthermore, the vortex-like fluid flow generated along the device edge effectively prevents precipitation due to excessive lipid concentration or contact with the wall.

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.

PM2.5 induces a senescent state in mouse AT2 cells.

PM2.5 is known to induce lung injury, but its toxic effects on lung regenerative machinery and the underlying mechanisms remain unknown. In this study, primary mouse alveolar type 2 (AT2) cells, considered stem cells in the gas-exchange barrier, were sorted using fluorescence-activated cell sorting. By developing microfluidic technology with constricted microchannels, we observed that both passage time and impedance opacities of mouse AT2 cells were reduced after PM2.5, indicating that PM2.5 induced a more deformable mechanical property and a higher membrane permeability. In vitro organoid cultures of primary mouse AT2 cells indicated that PM2.5 is able to impair the proliferative potential and self-renewal capacity of AT2 cells but does not affect AT1 differentiation. Furthermore, cell senescence biomarkers, p53 and γ-H2A.X at protein levels, P16ink4a and P21 at mRNA levels were increased in primary mouse AT2 cells after PM2.5 stimulations as shown by immunofluorescent staining and quantitative PCR analysis. Using several advanced single-cell technologies, this study sheds light on new mechanisms of the cytotoxic effects of atmospheric fine particulate matter on lung stem cell behavior.

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.

A Miniaturized Wireless Micropump Enabled by Confined Acoustic Streaming.

Miniaturization of health care, biomedical, and chemical systems is highly desirable for developing point-of-care testing (POCT) technologies. In system miniaturization, micropumps represent one of the major bottlenecks due to their undesirable pumping performance at such small sizes. Here, we developed a microelectromechanical system fabricated acoustic micropump based on an ultrahigh-frequency bulk acoustic wave resonator. The concept of an inner-boundary-confined acoustic jet was introduced to facilitate unidirectional flow. Benefitting from the high resonant frequency and confined acoustic streaming, the micropump reaches 32.620 kPa/cm3 (pressure/size) and 11.800 ml/min∙cm3 (flow rate/size), showing a 2-order-of-magnitude improvement in the energy transduction efficiency compared with the existing acoustic micropumps. As a proof of concept, the micropump was constructed as a wearable and wirelessly powered integrated drug delivery system with a size of only 9×9×9 mm3 and a weight of 1.16 g. It was demonstrated for ocular disease treatment through animal experimentation and a human pilot test. With superior pumping performance, miniaturized pump size, ultralow power consumption, and complementary metal-oxide-semiconductor compatibility, we expect it to be readily applied to various POCT applications including clinical diagnosis, prognosis, and drug delivery systems.

A Microflow Cytometer Enabled by Monolithic Integration of a Microreflector with an Acoustic Resonator.

Current microflow cytometers suffer from complicated fluidic integration and low fluorescence collection efficiency, resulting in reduced portability and sensitivity. Herein, we demonstrated a new flow cell design based on an on-chip monolithically integrated microreflector with a bulk acoustic wave resonator (MBAW). It enables simultaneous 3D particle focusing and fluorescence enhancement without using shear flow. Benefited by the on-chip microreflector, the captured fluorescence intensity was 1.8-fold greater than that of the Si substrate and 8.3-fold greater than that of the SiO2 substrate, greatly improving the detection sensitivity. Combined with the contactless acoustic streaming-based focusing, particle sensing with a coefficient of variation as low as 6.1% was achieved. We also demonstrated the difference between live and dead cells and performed a cell cycle assay using the as-developed microflow cytometry. This monolithic integrated MBAW provides a new type of opto-acoustofluidic system and has the potential to be a highly integrated, highly sensitive flow cytometer for applications such as in vitro diagnostics and point of care.

A novel ultrasonic-electric-based microsystem for the investigation of mechanical aging on lipid membranes.

Aging cells experience a gradual degeneration of their chemical and physical characteristics, resulting in the declining body functions that are commonly observed in old age. In this paper, we propose a novel microsystem that utilizes acoustics and electricity to investigate the effects of mechanical cyclic loading on cellular aging. Our study is the first to examine how mechanical loading influences the physicochemical characteristics of lipid bilayer at the subcellular level. By providing a new method for understanding the mechanism of cellular aging and aging-related diseases, our microsystem has significant implications for the development of treatments and therapies.Clinical Relevance- This ultrasonic-electric-based microsystem, as an in vitro model with sensitive quantitative capabilities, could have significant clinical implications in terms of understanding cellular responses to mechanical forces, elucidating the pathogenesis of aging-related diseases, and developing therapeutic strategies.

Recent Advances in Acoustofluidics for Point-of-Care Testing.

Point-of-care testing (POCT) has played important role in clinical diagnostics, environmental assessment, chemical and biological analyses, and food and chemical processing due to its faster turnaround compared to laboratory testing. Dedicated manipulations of solutions or particles are generally required to develop POCT technologies that achieve a "sample-in-answer-out" operation. With the development of micro- and nanotechnology, many tools have been developed for sample preparation, on-site analysis and solution manipulations (mixing, pumping, valving, etc.). Among these approaches, the use of acoustic waves to manipulate fluids and particles (named acoustofluidics) has been applied in many researches. This review focuses on the recent developments in acoustofluidics for POCT. It starts with the fundamentals of different acoustic manipulation techniques and then lists some of representative examples to highlight each method in practical POC applications. Looking toward the future, a compact, portable, highly integrated, low power, and biocompatible technique is anticipated to simultaneously achieve precise manipulation of small targets and multimodal manipulation in POC applications.

Pinch-off droplet generator using microscale gigahertz acoustics.

The generation and dispensing of microdroplets is a vital process in various fields such as biomedicine, medical diagnosis and chemistry. However, most methods still require the structures of nozzles or microchannels to assist droplet generation, which leads to limitations on system flexibility and restrictions on the size range of the generated droplets. In this paper, we propose a nozzle-free acoustic-based method for generating droplets using a gigahertz (GHz) bulk acoustic wave (BAW). Unlike most of the acoustofluidic approaches, the proposed method produces the droplet by pinching-off the liquid column generated by the acoustic body force at the oil-water interface. Benefitting from the focused acoustic energy and small footprint of the device, four orders of magnitude (ranging from 2 µm to 1800 µm) of droplet size could be produced by controlling the working time and power of the device. We also demonstrated cell encapsulation in the droplet and a high cell viability was achieved. The proposed acoustic-based droplet generation method exhibits capacity for generating droplets with a wide size range, versatility toward different viscosities, as well as biocompatibility for handling viable samples, which shows potential in miniaturization and scalability.

Rapid purification and enrichment of viral particles using self-propelled micromotors.

Virus infections remain one of the principal causes of morbidity and mortality worldwide. The current gold standard approach for diagnosing pathogens requires access to reverse transcription-polymerase chain reaction (RT-PCR) technology. However, separation and enrichment of the targets from complex and diluted samples remains a major challenge. In this work, we proposed a micromotor-based sample preparation concept for the efficient separation and concentration of target viral particles before PCR. The micromotors are functionalized with antibodies with a 3D polymer linker and are capable of self-propulsion by the catalytic generation of oxygen bubbles for selective and positive virus enrichment. This strategy significantly improves the enrichment efficiency and recovery rate of virus (up to 80% at 104 tu mL-1 in a 1 mL volume within just 6 min) without external mixing equipment. The method allows the Ct value in regular PCR tests to appear 6-7 cycles earlier and a detection limit of 1 tu mL-1 for the target virus from swap samples. A point-of-need test kit is designed based on the micromotors which can be readily applied to pretreat a large volume of samples.

Enhanced on-Chip modification and intracellular hydrogen peroxide detection via gigahertz acoustic streaming microfluidic platform.

Developing effective strategies for the flexible control of fluid is vital for microfluidic electrochemical biosensing. In this study, a gigahertz (GHz) acoustic streaming (AS) based sonoelectrochemical system was developed to realize an on-chip surface modification and sensitive hydrogen peroxide (H2O2) detection from living cells. The flexible and controlled fluid surrounding the electrochemical chip was optimized theoretically and applied in the sonoelectrochemical deposition of Au nanoparticles (AuNPs) first. Under the steady and fast flow stimulus of AS, AuNPs could be synthesized with a smaller and evener size distribution than the normal condition, allowing AuNPs to show an excellent peroxidase-like activity. Moreover, the AS also accelerated the mass transport of target molecules and improved the catalytic rate, leading to the enhancement of H2O2 detection, with an extremely low detection limit of 32 nM and a high sensitivity of 4.34 µA/ (mM·mm2). Finally, this system was successfully applied in tracking H2O2 release from different cell lines to distinguish the cancer cells from normal cells. This study innovatively integrated the surface modification and molecules detection process on a chip, and also proposed a simple but sensitive platform for microfluidic biosensing application.

Regulating Biomolecular Surface Interactions Using Tunable Acoustic Streaming.

Diffusion limitations and nonspecific surface absorption are great challenges for developing micro-/nanoscale affinity biosensors. There are very limited approaches that can solve these issues at the same time. Here, an acoustic streaming approach enabled by a gigahertz (GHz) resonator is presented to promote mass transfer of analytes through the jet mode and biofouling removal through the shear mode, which can be switched by tuning the deviation angle, α, between the resonator and the sensor. Simulations show that the jet mode (α ≤ 0) drives the analytes in the fluid toward the sensing surface, overcomes the diffusion limitation, and enhances the binding; while the shear mode (0 < α < π/4) provides a scouring action to remove the biofouling from the sensor. Experimental studies were performed by integrating this GHz resonator with optoelectronic sensing systems, where a 34-fold enhancement for the initial binding rate was obtained. Featuring high efficiency, controllability, and versatility, we believe that this GHz acoustic streaming approach holds promise for many kinds of biosensing systems as well as lab-on-chip systems.

Microchip gas chromatography column using magnetic beads coated with polydimethylsiloxane and metal organic frameworks.

Micro gas chromatography (µGC) using microfabricated silicon columns has been developed in response to the requirement for portable on-site gas analysis. Although different stationary phases have been developed, repeatable and reliable surface coatings in these rather small microcolumns remains a challenge. Herein, a new stationary phase coating strategy using magnetic beads (MBs) as carriers for micro column is presented. MBs modified with organopolysiloxane (MBs@OV-1) and a metal organic framework (MBs@HKUST-1) are deposited in on-chip microcolumns assisted with a magnetic field with an optimized modification process. MBs@OV-1 column showed a minimum HETP of 0.074 cm (1351 plates/m) of 62 cm/s. Mixtures of volatile organic compounds are successfully separated using MBs carried stationary phase which demonstrates that this technique has good chromatographic column efficiency. This method not only provides a novel coating process, washing and characterization of the stationary phases but also establishes a straightforward strategy for testing new absorbent materials for µGC systems.

Rational design of multivalent biosensor surfaces to enhance viral particle capture.

Viral particles bind to receptors through multivalent protein interactions. Such high avidity interactions on sensor surfaces are less studied. In this work, three polyelectrolytes that can form biosensing surfaces with different interfacial characteristics in probe density and spatial arrangement were designed. Quartz crystal microbalance, interferometry and atomic force microscopy were used to study their surface density and binding behaviors with proteins and virus particles. A multivalent adsorption kinetic model was developed to estimate the number of bonds from the viral particles bound to the polyelectrolyte surfaces. Experimental results show that the heterogeneous 3D surface with jagged forest-like structure enhances the virus capture ability by maximizing the multivalent interactions. As a proof of concept, specific coronavirus detection was achieved in spiked swab samples. These results indicate the importance of both probe density and their spatial arrangement on the sensing performance, which could be used as a guideline for rational biosensing surface design.

Tunable nanochannel resistive pulse sensing device using a novel multi-module self-assembly.

Nanochannel-based resistive pulse sensing (nano-RPS) system is widely used for the high-sensitive measurement and characterization of nanoscale biological particles and biomolecules due to its high surface to volume ratio. However, the geometric dimensions and surface properties of nanochannel are usually fixed, which limit the detections within particular ranges or types of nanoparticles. In order to improve the flexibility of nano-RPS system, it is of great significance to develop nanochannels with tunable dimensions and surface properties. In this work, we proposed a novel multi-module self-assembly (MS) strategy which allows to shrink the geometric dimensions and tune surface properties of the nanochannels simultaneously. The MS-tuned nano-RPS device exhibits an enhanced signal-to-noise ratio (SNR) for nanoparticle detections after shrunk the geometric dimensions by MS strategy. Meanwhile, by tuning the surface charge, an enhanced resolution for viral particles detection was achieved with the MS-tuned nano-RPS devices by analyzing the variation of pulse width due the tuned surface charge. The proposed MS strategy is versatile for various types of surface materials and can be potentially applied for nanoscale surface reconfiguration in various nanofluidic devices.

Ultrasensitive Linear Capacitive Pressure Sensor with Wrinkled Microstructures for Tactile Perception.

Ultrasensitive flexible pressure sensors with excellent linearity are essential for achieving tactile perception. Although microstructured dielectrics have endowed capacitive sensors with ultrahigh sensitivity, the compromise of sensitivity with increasing pressure is an issue yet to be resolved. Herein, a spontaneously wrinkled MWCNT/PDMS dielectric layer is proposed to realize the excellent sensitivity and linearity of capacitive sensors for tactile perception. The synergistic effect of a high dielectric constant and wrinkled microstructures enables the sensor to exhibit linearity up to 21 kPa with a sensitivity of 1.448 kPa-1 and a detection limit of 0.2 Pa. Owing to these merits, the sensor monitors subtle physiological signals such as various arterial pulses and respiration. This sensor is further integrated into a fully multimaterial 3D-printed soft pneumatic finger to realize material hardness perception. Eight materials with different hardness values are successfully discriminated, and the capacitance of the sensor varies linearly (R2 > 0.975) with increasing hardness. Moreover, the sensitivity to the material hardness can be tuned by controlling the inflation pressure of the soft finger. As a proof of concept, the finger is used to discriminate pork fats with different hardness, paving the way for hardness discrimination in clinical palpation.

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.