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Acoustic tweezers have gained substantial interest in biology, engineering, and materials science for their label-free, precise, contactless, and programmable manipulation of small objects. However, acoustic tweezers cannot independently manipulate multiple microparticles simultaneously. This study introduces acousto-dielectric tweezers capable of independently manipulating multiple microparticles and precise control over intercellular distances and cyclical cell pairing and separation for detailed cell-cell interaction analysis. Our acousto-dielectric tweezers leverage the competition between acoustic radiation forces, generated by standing surface acoustic waves (SAWs), and dielectrophoretic (DEP) forces, induced by gradient electric fields. Modulating these fields allows for the precise positioning of individual microparticles at points where acoustic radiation and DEP forces are in equilibrium. This mechanism enables the simultaneous movement of multiple microparticles along specified paths as well as cyclical cell pairing and separation. We anticipate our acousto-dielectric tweezers to have enormous potential in colloidal assembly, cell-cell interaction studies, disease diagnostics, and tissue engineering.
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Pinzas Ópticas , Acústica , HumanosRESUMEN
Hydrogel microspheres are biocompatible materials widely used in biological and medical fields. Emulsification and stirring are the commonly used methods to prepare hydrogels. However, the size distribution is considerably wide, the monodispersity and the mechanical intensity are poor, and the stable operation conditions are comparatively narrow to meet some sophisticated applications. In this paper, a T-shaped stepwise microchannel combined with a simple side microchannel structure is developed to explore the liquid-liquid dispersion mechanism, interfacial evolution behavior, satellite droplet formation mechanism and separation, and the eventual successful synthesis of dextran hydrogel microspheres. The effect of the operation parameters on droplet and microsphere size is comprehensively studied. The flow pattern and the stable operation condition range are given, and mathematical prediction models are developed under three different flow regimes for droplet size prediction. Based on the stable operating conditions, a microdroplet-based method combined with UV light curing is developed to synthesize the dextran hydrogel microsphere. The highly uniform and monodispersed dextran microspheres with good mechanical intensity are synthesized in the developed microfluidic platform. The size of the microsphere could be tuned from 50 to 300 µm with a capillary number in the range of 0.006-0.742. This work not only provides a facile method for functional polymeric microsphere preparation but also offers important design guidelines for the development of a robust microreactor.
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SERS measurements for monitoring bactericides in dairy products are highly desired for food safety problems. However, the complicated preparation process of SERS substrates greatly impedes the promotion of SERS. Here, we propose acoustofluidic one-step synthesis of Ag nanoparticles on paper substrates for SERS detection. Our method is economical, fast, simple, and eco-friendly. We adopted laser cutting to cut out appropriate paper shapes, and aldehydes were simultaneously produced at the cutting edge in the pyrolysis of cellulose by laser which were leveraged as the reducing reagent. In the synthesis, only 5 µL of Ag precursor was added to complete the reaction, and no reducing agent was used. Our recently developed acoustofluidic device was employed to intensely mix Ag+ ions and aldehydes and spread the reduced Ag nanoparticles over the substrate. The SERS substrate was fabricated in 1 step and 3 min. The standard R6G solution measurement demonstrated the excellent signal and prominent uniformity of the fabricated SERS substrates. SERS detection of the safe concentration of three bactericides, including tetracycline hydrochloride, thiabendazole, and malachite green, from food samples can be achieved using fabricated substrates. We take the least cost, time, reagents, and steps to fabricate the SERS substrate with satisfying performance. Our work has an extraodinary meaning for the green preparation and large-scale application of SERS.
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Antibacterianos , Nanopartículas del Metal , Papel , Plata , Espectrometría Raman , Plata/química , Nanopartículas del Metal/química , Antibacterianos/análisis , Propiedades de Superficie , Tetraciclina/análisis , Colorantes de Rosanilina/análisis , Colorantes de Rosanilina/química , Tiabendazol/análisis , Tamaño de la PartículaRESUMEN
Manipulating objects with acoustics has been developed for hundreds of years since Chladni patterns in gaseous environments were exhibited. In recent decades, acoustic manipulation in microfluidics, known as acoustofluidics, has rapidly thrived and many sophisticated technologies were born. However, the basic background motion of particles under acoustic excitation is usually neglected and the classical Chladni patterns haven't been reproduced in an aqueous environment. In this study, we investigated the basic mechanism and the motion of suspended particles and sinking particles in a plain microchamber under low-frequency excitation (3-5 kHz). The mechanisms were clearly distinguished by comparing the differences among colored fluids, suspended particles, and sinking particles. The suspended particles rotated around the antinode with a speed up to 55.1 µm s-1 at 100 Vpp by the acoustic streaming and they approached each other by the secondary acoustic radiation force. The sinking particles concentrated at the node with a speed up to 22.3 µm s-1 at 100 Vpp by bouncing on the vibrating surface and the primary acoustic radiation force. We have reproduced the classical standard/inverse Chladni patterns in an aqueous environment for the first time, and they were leveraged to separate SiO2 particles with different sizes. The big particles with an average diameter of 9.68 µm were concentrated at the node while the small particles with an average diameter of 2.72 µm were collected at the antinode within 2 min. These results not only provide insightful perspectives of basic mechanisms, but also open up new possibilities for advanced acoustic tweezers.
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Surface acoustic wave (SAW)-enabled acoustofluidic technologies have recently atttracted increasing attention for applications in biology, chemistry, biophysics, and medicine. Most SAW acoustofluidic devices generate acoustic energy which is then transmitted into custom microfabricated polymer-based channels. There are limited studies on delivering this acoustic energy into convenient commercially-available glass tubes for manipulating particles and fluids. Herein, we have constructed a capillary-based SAW acoustofluidic device for multifunctional fluidic and particle manipulation. This device integrates a converging interdigitated transducer to generate focused SAWs on a piezoelectric chip, as well as a glass capillary that transports particles and fluids. To understand the actuation mechanisms underlying this device, we performed finite element simulations by considering piezoelectric, solid mechanic, and pressure acoustic physics. This experimental study shows that the capillary-based SAW acoustofluidic device can perform multiple functions including enriching particles, patterning particles, transporting particles and fluids, as well as generating droplets with controlled sizes. Given the usefulness of these functions, we expect that this acoustofluidic device can be useful in applications such as pharmaceutical manufacturing, biofabrication, and bioanalysis.
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Compared with the conventional emulsification method, droplets generated within microfluidic devices exhibit distinct advantages such as precise control of fluids, exceptional monodispersity, uniform morphology, flexible manipulation, and narrow size distribution. These inherent benefits, including intrinsic safety, excellent heat and mass transfer capabilities, and large surface-to-volume ratio, have led to the widespread applications of droplet-based microfluidics across diverse fields, encompassing chemical engineering, particle synthesis, biological detection, diagnostics, emulsion preparation, and pharmaceuticals. However, despite its promising potential for versatile applications, the practical utilization of this technology in commercial and industrial is extremely limited to the inherently low production rates achievable within a single microchannel. Over the past two decades, droplet-based microfluidics has evolved significantly, considerably transitioning from a proof-of-concept stage to industrialization. And now there is a growing trend towards translating academic research into commercial and industrial applications, primarily driven by the burgeoning demands of various fields. This paper comprehensively reviews recent advancements in droplet-based microfluidics, covering the fundamental working principles and the critical aspect of scale-up integration from working principles to scale-up integration. Based on the existing scale-up strategies, the paper also outlines the future research directions, identifies the potential opportunities, and addresses the typical unsolved challenges.
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Microfluidic paper-based analytical devices (µPADs) feature an economic and sensitive nature, while acoustofluidics displays contactless and versatile virtue, and both of them gained tremendous interest in the past decades. Integrating µPADs with acoustofluidic techniques provides great potential to overcome the inherent shortcomings and make appealing achievements. Here, we present acoustofluidics-assisted multifunctional paper-based analytical devices that leverage bulk acoustic waves to realize multiple applications on paper substrates, including uniform colorimetric detection, microparticle/cell enrichment, fluorescence amplification, homogeneous mixing, and nanomaterial synthesis. The glucose detection in the range of 5-15 mM was conducted to perform uniform colorimetric detection. Various types (brass powder, copper powder, diamond powder, and yeast cells) and sizes (5-200 µm) of solid particles and biological cells can be enriched on paper in a few seconds or minutes; thus, fluorescence amplification by 3 times was realized with the enrichment. The high-throughput and homogeneous mixing of two fluids can be achieved, and based on the mixing, nanomaterials (ZnO nanosheets) were synthesized on paper. We analyzed the underlying mechanisms of these applications in the devices, which are attributed to Faraday waves and Chladni patterns. With their simple fabrication and prominent effectiveness, the devices open up new possibilities for paper-based microfluidic devices.
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The intrinsic biophysical properties of cells, such as mechanical, acoustic, and electrical properties, are valuable indicators of a cell's function and state. However, traditional single-cell biophysical characterization methods are hindered by limited measurable properties, time-consuming procedures, and complex system setups. This study presents acousto-dielectric tweezers that leverage the balance between controllable acoustophoretic and dielectrophoretic forces applied on cells through surface acoustic waves and alternating current electric fields, respectively. Particularly, the balanced acoustophoretic and dielectrophoretic forces can trap cells at equilibrium positions independent of the cell size to differentiate between various cell-intrinsic mechanical, acoustic, and electrical properties. Experimental results show our mechanism has the potential for applications in single-cell analysis, size-insensitive cell separation, and cell phenotyping, which are all primarily based on cells' intrinsic biophysical properties. Our results also show the measured equilibrium position of a cell can inversely determine multiple biophysical properties, including membrane capacitance, cytoplasm conductivity, and acoustic contrast factor. With these features, our acousto-dielectric tweezing mechanism is a valuable addition to the resources available for biophysical property-based biological and medical research.
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Técnicas Biosensibles , Sonido , Citoplasma , Conductividad Eléctrica , AcústicaRESUMEN
High-precision isolation of small extracellular vesicles (sEVs) from biofluids is essential toward developing next-generation liquid biopsies and regenerative therapies. However, current methods of sEV separation require specialized equipment and time-consuming protocols and have difficulties producing highly pure subpopulations of sEVs. Here, we present Acoustic Nanoscale Separation via Wave-pillar Excitation Resonance (ANSWER), which allows single-step, rapid (<10 min), high-purity (>96% small exosomes, >80% exomeres) fractionation of sEV subpopulations from biofluids without the need for any sample preprocessing. Particles are iteratively deflected in a size-selective manner via an excitation resonance. This previously unidentified phenomenon generates patterns of virtual, tunable, pillar-like acoustic field in a fluid using surface acoustic waves. Highly precise sEV fractionation without the need for sample preprocessing or complex nanofabrication methods has been demonstrated using ANSWER, showing potential as a powerful tool that will enable more in-depth studies into the complexity, heterogeneity, and functionality of sEV subpopulations.
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Nanocarrier and exosome encapsulation has been found to significantly increase the efficacy of targeted drug delivery while also minimizing unwanted side effects. However, the development of exosome-encapsulated drug nanocarriers is limited by low drug loading efficiencies and/or complex, time-consuming drug loading processes. Herein, we have developed an acoustofluidic device that simultaneously performs both drug loading and exosome encapsulation. By synergistically leveraging the acoustic radiation force, acoustic microstreaming, and shear stresses in a rotating droplet, the concentration, and fusion of exosomes, drugs, and porous silica nanoparticles is achieved. The final product consists of drug-loaded silica nanocarriers that are encased within an exosomal membrane. The drug loading efficiency is significantly improved, with nearly 30% of the free drug (e.g., doxorubicin) molecules loaded into the nanocarriers. Furthermore, this acoustofluidic drug loading system circumvents the need for complex chemical modification, allowing drug loading and encapsulation to be completed within a matter of minutes. These exosome-encapsulated nanocarriers exhibit excellent efficiency in intracellular transport and are capable of significantly inhibiting tumor cell proliferation. By utilizing physical forces to rapidly generate hybrid nanocarriers, this acoustofluidic drug loading platform wields the potential to significantly impact innovation in both drug delivery research and applications.
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Acoustic black holes offer superior capabilities for slowing down and trapping acoustic waves for various applications such as metastructures, energy harvesting, and vibration and noise control. However, no studies have considered the linear and nonlinear effects of acoustic black holes on micro/nanoparticles in fluids. This study presents acoustofluidic black holes (AFBHs) that leverage controlled interactions between AFBH-trapped acoustic wave energy and particles in droplets to enable versatile particle manipulation functionalities, such as translation, concentration, and patterning of particles. We investigated the AFBH-enabled wave energy trapping and wavelength shrinking effects, as well as the trapped wave energy-induced acoustic radiation forces on particles and acoustic streaming in droplets. This study not only fills the gap between the emerging fields of acoustofluidics and acoustic black holes but also leads to a class of AFBH-based in-droplet particle manipulation toolsets with great potential for many applications, such as biosensing, point-of-care testing, and drug screening.
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Microrobots can operate in tiny areas that traditional bulk robots cannot reach. The combination of acoustic actuation with microrobots extensively expands the application areas of microrobots due to their desirable miniaturization, flexibility, and biocompatibility features. Herein, an overview of the research and development of acoustics-actuated microrobots is provided. We first introduce the currently established manufacturing methods (3D printing and photolithography). Then, according to their different working principles, we divide acoustics-actuated microrobots into three categories including bubble propulsion, sharp-edge propulsion, and in-situ microrotor. Next, we summarize their established applications from targeted drug delivery to microfluidics operation to microsurgery. Finally, we illustrate current challenges and future perspectives to guide research in this field. This work not only gives a comprehensive overview of the latest technology of acoustics-actuated microrobots, but also provides an in-depth understanding of acoustic actuation for inspiring the next generation of advanced robotic devices.
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Alzheimer's disease (AD) is a progressive and irreversible neurodegenerative brain disorder that affects tens of millions of older adults worldwide and has significant economic and societal impacts. Despite its prevalence and severity, early diagnosis of AD remains a considerable challenge. Here we report an integrated acoustofluidics-based diagnostic system (ADx), which combines triple functions of acoustics, microfluidics, and orthogonal biosensors for clinically accurate, sensitive, and rapid detection of AD biomarkers from human plasma. We design and fabricate a surface acoustic wave-based acoustofluidic separation device to isolate and purify AD biomarkers to increase the signal-to-noise ratio. Multimodal biosensors within the integrated ADx are fabricated by in-situ patterning of the ZnO nanorod array and deposition of Ag nanoparticles onto the ZnO nanorods for surface-enhanced Raman scattering (SERS) and electrochemical immunosensors. We obtain the label-free detections of SERS and electrochemical immunoassay of clinical plasma samples from AD patients and healthy controls with high sensitivity and specificity. We believe that this efficient integration provides promising solutions for the early diagnosis of AD.
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Enfermedad de Alzheimer , Técnicas Biosensibles , Nanopartículas del Metal , Anciano , Enfermedad de Alzheimer/diagnóstico , Humanos , Inmunoensayo , Plata , Espectrometría RamanRESUMEN
The ability to precisely manipulate nano-objects on a large scale can enable the fabrication of materials and devices with tunable optical, electromagnetic, and mechanical properties. However, the dynamic, parallel manipulation of nanoscale colloids and materials remains a significant challenge. Here, we demonstrate acoustoelectronic nanotweezers, which combine the precision and robustness afforded by electronic tweezers with versatility and large-field dynamic control granted by acoustic tweezing techniques, to enable the massively parallel manipulation of sub-100 nm objects with excellent versatility and controllability. Using this approach, we demonstrated the complex patterning of various nanoparticles (e.g., DNAs, exosomes, ~3 nm graphene flakes, ~6 nm quantum dots, ~3.5 nm proteins, and ~1.4 nm dextran), fabricated macroscopic materials with nano-textures, and performed high-resolution, single nanoparticle manipulation. Various nanomanipulation functions, including transportation, concentration, orientation, pattern-overlaying, and sorting, have also been achieved using a simple device configuration. Altogether, acoustoelectronic nanotweezers overcome existing limitations in nano-manipulation and hold great potential for a variety of applications in the fields of electronics, optics, condensed matter physics, metamaterials, and biomedicine.
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Electrónica/métodos , Nanoestructuras/química , Nanotecnología/métodos , Pinzas Ópticas , Acústica , Coloides/química , ADN/química , Exosomas/química , Nanopartículas del Metal/química , Nanotubos de Carbono , Tamaño de la Partícula , Reproducibilidad de los ResultadosRESUMEN
BACKGROUND: Hyperthermia is one of the promising cancer treatment strategies enabled by local heating with the use of tumor-targeting magnetic nanoparticles (MNP) under a non-invasive magnetic field. However, one of the remaining challenges is how to achieve therapeutic levels of heat (without causing damages to regular tissues) in tumors that cannot be effectively treated with anti-tumor drug delivery. RESULTS: In this work, we report a facile method to fabricate magnetic nanorods for hyperthermia by one-step wet chemistry synthesis using 3-Aminopropyltrimethoxysilane (APTMS) as the shape-controlling agent and ferric and ferrous ions as precursors. By adjusting the concentration of APTMS, hydrothermal reaction time, ratios of ferric to ferrous ions, magnetic nanorods with aspect ratios ranging from 4.4 to 7.6 have been produced. At the clinically recommended field strength of 300 Oe (or less) and the frequency of 184 kHz, the specific absorption rate (SAR) of these nanorods is approximately 50 % higher than that of commercial Bionized NanoFerrite particles. CONCLUSIONS: This increase in SAR, especially at low field strengths, is crucial for treating deep tumors, such as pancreatic and rectal cancers, by avoiding the generation of harmful eddy current heating in normal tissues.
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Antineoplásicos/farmacología , Hipertermia/tratamiento farmacológico , Magnetismo , Nanopartículas/uso terapéutico , Nanotubos/química , Compuestos Férricos/uso terapéutico , Calefacción , Calor , Humanos , Hipertermia Inducida/métodos , Campos Magnéticos , Neoplasias/tratamiento farmacológicoRESUMEN
Opioid abuse is a significant public health problem. Over two million Americans have some form of addiction to opioids; however, despite governmental programs established to treat overdoses and restrict opioid distribution, there are still few screening tools that are quantitative, portable and easy to use for high-throughput mapping and monitoring this ongoing crisis. In this paper, we demonstrated a plasmonic zinc oxide (ZnO) arrays-on-silicon sensor for the label-free detection of opioids through surface-enhanced Raman spectroscopy (SERS), and evaluated the chips' opioid sensing performance. Specifically, we tested our device with oxycodone, a potent and commonly abused opioid, dissolved in methanol and blood serum as a proof-of-concept study. Ag particles were in situ patterned onto the ZnO array to form the completed sensing platform. The resulting Ag@ZnO arrays were characterized using Scanning Electron Microscopy (SEM), Energy Dispersive X-ray Analysis (EDS), and element mapping. In addition, the enhanced electric field induced by the localized surface plasmonic resonance at the Ag particle decorated ZnO is simulated using COMSOL. Opioid-containing samples at varying concentrations, from 900 µg mL-1 to 90 ng mL-1 were tested using SERS to characterize the chip's accuracy and sensitivity. We demonstrated that the sensor can reliably detect opioid concentrations as low as 90 ng mL-1 with great accuracy and sensitivity even spiked into blood serum. The chips could provide a cost-effective, high-throughput method for detecting opiate oxycodone, thereby providing a powerful tool to monitor and control the emerging public health threats.
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Piezoelectric nanomaterial-polymer composites represent a unique paradigm for making flexible energy harvesting and sensing devices with enhanced devices' performance. In this work, we studied various metal doped ZnO nanostructures, fabricated and characterized ZnO nanoparticle-PVDF composite thin film, and demonstrated both enhanced energy generation and motion sensing capabilities. Specifically, a series of flexible piezoelectric nanogenerators (PENGs) were designed based on these piezoelectric composite thin films. The voltage output from cobalt (Co), sodium (Na), silver (Ag), and lithium (Li) doped ZnO-PVDF composite as well as pure ZnO-PVDF samples were individually studied and compared. Under the same experimental conditions, the Li-ZnO based device produces the largest peak-to-peak voltage (3.43 Vpp) which is about 9 times of that of the pure ZnO based device, where Co-ZnO, Na-ZnO and Ag-ZnO are 1.2, 4.9 and 5.4 times, respectively. In addition, the effect of doping ratio of Li-ZnO is studied, and we found that 5% is the best doping ratio in terms of output voltage. Finally, we demonstrated that the energy harvested by the device from finger tapping at ~2 Hz can charge a capacitor with a large output power density of 0.45 W/cm3 and light up an ultraviolet (UV) light-emitting diode (LED). We also showed the device as a flexible wearable motion sensor, where different hand gestures were detected by the device with distinctive output voltage amplitudes and patterns.
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Acoustofluidics, the fusion of acoustics and microfluidic techniques, has recently seen increased research attention across multiple disciplines due in part to its capabilities in contactless, biocompatible, and precise manipulation of micro-/nano-objects. Herein, a bimodal signal amplification platform which relies on acoustofluidics-induced enrichment of nanoparticles is introduced. The dual-function biosensor can perform sensitive immunofluorescent or surface-enhanced Raman spectroscopy (SERS) detection. The platform functions by using surface acoustic waves to concentrate nanoparticles at either the center or perimeter of a glass capillary; the concentration location is adjusted simply by varying the input frequency. The immunofluorescence assay is achieved by concentrating fluorescent analytes and functionalized nanoparticles at the center of the microchannel, thereby improving the visibility of the fluorescent output. By modifying the inner wall of the glass capillary with plasmonic Ag nanoparticle-deposited ZnO nanorod arrays and focusing analytes toward the perimeter of the microchannel, SERS sensing using the same device setup is achieved. Nanosized exosomes are used as a proof-of-concept to validate the performance of the acoustofluidic bimodal biosensor. With its sample-enrichment functionality, bimodal sensing, short processing time, and minute sample consumption, the acoustofluidic chip holds great potential for the development of lab-on-a-chip based analysis systems in many real-world applications.
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Técnicas Biosensibles , Nanopartículas del Metal , Dispositivos Laboratorio en un Chip , Plata , Espectrometría RamanRESUMEN
Metal-organic framework (MOF)-based drug delivery nanosystems with both precise drug release and multidrug codelivery capabilities have emerged as promising candidates for cancer treatment. However, challenges are posed by the limited number of suitable payload types, uncontrollable drug leakage, and lack of chemical groups for postmodification. To overcome those challenges, we developed a core-shell nanocomposite composed of zeolitic imidazolate framework-90 (ZIF-90) coated with spermine-modified acetalated dextran (SAD) by a facile microfluidics-based nanoprecipitation method. This nanocomposite could serve as a multidrug storage reservoir for the loading of two drugs with distinct properties, where the hydrophilic doxorubicin (DOX) was coordinately attached to the ZIF-90 framework, and hydrophobic photosensitizer IR780 was loaded into the SAD shell, enabling the combination therapy of photodynamic treatment with chemotherapy. Meanwhile, equipping ZIF-90 with a SAD shell not only substantially improved the pH-responsive drug release of ZIF-90 but also enabled the postformation conjugation of ZIF-90 with hyaluronic acid for specific CD44 recognition, thereby facilitating precise drug delivery to CD44-overexpressed tumor. Such a simple microfluidics-based strategy can efficiently overcome the limitations of solely MOF-based DDSs and greatly extend the flexibility of MOF biomedical applications.
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Antibióticos Antineoplásicos/farmacología , Doxorrubicina/farmacología , Estructuras Metalorgánicas/química , Técnicas Analíticas Microfluídicas , Nanopartículas/química , Animales , Antibióticos Antineoplásicos/química , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Doxorrubicina/química , Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Células HeLa , Humanos , Imidazoles/química , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Estructura Molecular , Neoplasias Experimentales/diagnóstico por imagen , Neoplasias Experimentales/tratamiento farmacológico , Tamaño de la Partícula , Fotoquimioterapia , Propiedades de Superficie , Zeolitas/químicaRESUMEN
Controllable enrichment of micro/nanoscale objects plays a significant role in many biomedical and biochemical applications, such as increasing the detection sensitivity of assays, or improving the structures of bio-engineered tissues. However, few techniques can perform concentrations of micro/nano objects in multi-well plates, a very common laboratory vessel. In this work, we develop an acoustofluidic multi-well plate, which adopts an array of simple, low-cost and commercially available ring-shaped piezoelectric transducers for rapid and robust enrichment of micro/nanoscale particles/cells in each well of the plate. The enrichment mechanism is validated and characterized through both numerical simulations and experiments. We observe that the ring-shaped piezoelectric transducer can generate circular standing flexural waves in the substrate of each well, and that the vibrations can induce acoustic streaming near the interface between the substrate and a fluid droplet placed within the well; this streaming can drive micro/nanoscale objects to the center of the droplet for enrichment. Moreover, the acoustofluidic multi-well plate can realize simultaneous and consistent enrichment of biological cells in each well of the plate. With merits such as simplicity, controllability, low cost, and excellent compatibility with other downstream analysis tools, the developed acoustofluidic multi-well plate could be a versatile tool for many applications such as micro/nano fabrication, self-assembly, biomedical/biochemical sensing, and tissue engineering.