Tony Jun Huang

Dynamic acoustofluidics enables precise, contact-free manipulation of particles, colloids, and cells and shows great potential for applications in physics, materials science, and life sciences. However, existing strategies struggle to realize contrast-based selective manipulation primarily because the pressure fields are time invariant. Here, we introduce a space-time acoustofluidic tweezer (STAT) that uses frequency detuning-induced pseudo-space-time modulation of standing surface acoustic waves to enable dynamic, contrast-dependent control of microparticles and cells. Experiments and simulations show that, under STAT manipulation, positive (PACP) and negative (NACP) acoustic contrast particles can undergo low-frequency, shear- and longitudinal-like harmonic motions, respectively. Under certain driving conditions, NACPs can be selectively guided along programmed paths, whereas PACPs remain stably patterned. Overall, STAT offers a gentle, biocompatible way to selectively drive oscillation, transport, and sorting among particles and cells of different acoustic contrasts, broadening the capabilities of acoustofluidic systems for biomedical applications.

Proteolysis-targeting chimeras (PROTACs) have emerged as a promising molecular approach for degrading undruggable proteins and for overcoming drug resistance in cancer therapy. However, their clinical translation remains limited by challenges such as poor cell membrane permeability, limited intracellular uptake, and potential off-target toxicity. To overcome these barriers, we developed Sonoporation-assisted Precise Intracellular Nanodelivery (SonoPIN), an ultrasound-driven, aptamer-guided microbubble system that enables rapid delivery of therapeutic molecules with cell selectivity. By leveraging aptamer-conjugated microbubbles and ultrasound-induced sonoporation, SonoPIN transiently permeabilizes the membranes of target cells, while leaving nontarget cells undisturbed. Using BRD4, a well-characterized oncogenic transcriptional coactivator and validated PROTAC target critically involved in cancer cell survival, as a model system, we demonstrate that SonoPIN facilitates highly efficient intracellular delivery of fluorescently labeled PROTACs. SonoPIN achieves a sevenfold increase in intracellular fluorescence after 60 s of ultrasound stimulation, resulting in a 70% reduction in BRD4 protein levels specifically in cancer cells. Importantly, BRD4 degradation is undetectable in noncancerous cells. Consequently, approximately 50% of the targeted cancer cells undergo apoptosis while nontarget cells retain more than 99% viability, underscoring the high selectivity of the SonoPIN system. Our study indicates that SonoPIN represents an innovative, noninvasive delivery platform for PROTAC therapeutics, offering a rapid and precise approach for targeted drug delivery in cancer treatment.

Microrobotic systems offer significant potential for precision medicine by enabling minimally invasive interventions in complex fluidic environments. However, effective operation in these settings requires actuators capable of more than simple linear or rotational motion, often necessitating programmable changes in both direction and shape. This remains a major challenge due to fundamental constraints in the design and control of microscale actuators, particularly in acoustic systems. Here, we introduce engineered cilia for hybrid operations microrobots, a class of acoustic microrobots that use geometry-tuned cilia and resonance-induced forces to execute complex motions such as bidirectional bending, controllable rotation, and adaptive morphing. The microrobots design is driven by a self-augmenting machine learning framework integrated with finite element analysis, enabling rapid prediction and optimization of geometry-resonance relationships across design space. This approach achieves >10⁵-fold reduction in prediction time and over 20-fold in memory savings, while maintaining >90% accuracy in peak amplitude and >98% in resonance frequency. Compliant mechanism strategies further expand the mechanical versatility of the microrobots, enabling programmable shape transformations tailored to specific tasks. These advances establish acoustic-driven microrobots as a scalable and efficient platform for intelligent microrobotic actuation in biomedical and microfluidic applications.

Guided elastic waves are a truly cross-disciplinary key enabling technology. For more than five decades, surface acoustic wave (SAW) and bulk acoustic wave devices find widespread applications. Nowadays, different types of guided elastic waves cover the wide spectrum of applications spanning from quantum technologies to the life sciences, from controlling single excitations to macroscopic collective states in condensed matter. Six years after the first 2019 SAW roadmap, we believe it is time to make a step back and take a fresh look at the status of the field and its future challenges. Since the first roadmap in 2019, the spectrum clearly expanded and this new edition presents a current snapshot of the status of this vibrant field and prospects for potential future developments.

Mechanomodulation, the process of altering cellular behavior through applied mechanical forces, plays a critical role in physiological processes and has substantial implications for cancer therapy, immunology, and drug development. However, precise and efficient stimulation of nonadherent cells remains a major challenge, limiting the investigation of mechanotransduction pathways and the development of targeted therapeutics. Here, we developed an acoustofluidic platform named Suspension-cell Targeted Response to Excitation via Acoustofluidic Mechanomodulation (STREAM) to enable precise, high-throughput stimulation of suspension cells. STREAM accomplishes this using 101.14-megahertz high-frequency surface acoustic waves to deliver controlled mechanical stimulation at a throughput of 500,000 cells per minute. STREAM modulates intracellular calcium ion (Ca 2+ ) signaling by activating mechanosensitive ion channels, triggering mitochondrial membrane disruption and tunable K562 leukemia cell apoptosis rates from 5.15 to 47.1%. STREAM provides a scalable, precise tool for studying mechanotransduction in suspension cells, with broad applications in cancer research, immunotherapy, and high-throughput drug screening.

The code include the ML model to predict cilia resonance peak features and compared to FEA results.

biosensors, these systems provide unmatched benefits in sensitivity, speed, portability, and immediate monitoring, thereby transforming diagnostics in human and animal health, environmental sensing, and point-of-care testing. In this review, we provide a comprehensive overview of integrated microfluidics with biosensors, highlighting the synergistic interplay between these two complementary fields and their various biomedical applications. We begin by examining different microfluidic technologies, including 3D dynamic cell culture systems, inertial microfluidic separation, acoustofluidics, dielectrophoresis, optofluidics, and immunoassays. Next, we discuss integrated microfluidic systems that incorporate various biosensor technologies, including electrochemical, electrophysiological, plasmonic, Raman, and quantum sensors. These are designed to detect and analyze DNA, RNA, proteins, exosomes, cells, and small organisms, covering a size range from nanometers to millimeters. Additionally, we discuss the wide range of applications for integrated microfluidic biosensors and examine significant challenges and future opportunities that will influence their ongoing development and practical use. Finally, we highlight successful commercial products developed with integrated microfluidic technologies.

Gradient biomaterials that exhibit spatially varying physical, chemical, or biological properties can be used in various applications such as tissue engineering, organoid development, mechanobiology, and spatially controlled drug delivery. However, current fabrication methods often suffer from limited gradient precision, restricted material compatibility, and poor reproducibility. Here, we introduced gradient regulation via acoustofluidic dynamic engineering (GRADE), a programmable system to generate high-fidelity gradient biomaterials across different material systems. By incorporating focused interdigital transducers with the pulsed surface acoustic wave actuation, GRADE can achieve tunable and directional acoustic streaming (0 to 22 millimeters per second), which allows accurate regulation of the gradient magnitude and length. Its open microchannel design enables nondestructive extraction of centimeter-scale gradients and supports device reuse, enhancing practicality and scalability. In contrast to magnetic or electrospinning techniques that are limited to specific material types, the GRADE approach supports composition-independent fluid manipulation of a diverse group of biomaterials and cross-linking methods, thus providing greater versatility and translational potential. Furthermore, we demonstrate stiffness-dependent mechanosensation in stem cells cultured on customized gradient substrates, which validates the platform's usability. The experimental results show that GRADE has the ability to uncover mechanobiological responses in physiologically relevant contexts. All these results establish GRADE as a powerful and versatile platform for gradient biomaterial fabrication. It shows broad potential to advance both fundamental research and translational biomedical applications.

Gradient biomaterials that exhibit spatially varying physical, chemical, or biological properties can be used in various applications such as tissue engineering, organoid development, mechanobiology, and spatially controlled drug delivery. However, current fabrication methods often suffer from limited gradient precision, restricted material compatibility, and poor reproducibility. Here, we introduced gradient regulation via acoustofluidic dynamic engineering (GRADE), a programmable system to generate high-fidelity gradient biomaterials across different material systems. By incorporating focused interdigital transducers with the pulsed surface acoustic wave actuation, GRADE can achieve tunable and directional acoustic streaming (0 to 22 millimeters per second), which allows accurate regulation of the gradient magnitude and length. Its open microchannel design enables nondestructive extraction of centimeter-scale gradients and supports device reuse, enhancing practicality and scalability. In contrast to magnetic or electrospinning techniques that are limited to specific material types, the GRADE approach supports composition-independent fluid manipulation of a diverse group of biomaterials and cross-linking methods, thus providing greater versatility and translational potential. Furthermore, we demonstrate stiffness-dependent mechanosensation in stem cells cultured on customized gradient substrates, which validates the platform’s usability. The experimental results show that GRADE has the ability to uncover mechanobiological responses in physiologically relevant contexts. All these results establish GRADE as a powerful and versatile platform for gradient biomaterial fabrication. It shows broad potential to advance both fundamental research and translational biomedical applications.

Liquid biopsies that analyze the molecular content of extracellular vesicles and particles (EVPs) are a formidable opportunity for early and late-stage cancer diagnosis. This study explores the potential of two advanced technologies: Bessel beam excitation separation technology (BEST) and multiparametric biochip assay (MBA), to track single EVP cargo from organs of pathology into biofluids such as plasma and saliva. Using gastric cancer (GC) as a disease model, we conducted high-throughput, multiparametric analyses of EVPs derived from plasma, saliva, and tissue samples from GC patients. Our findings demonstrate the feasibility of these techniques in isolating and characterizing EVPs, revealing consistent EVP morphology and size across biofluids. Furthermore, differential expression patterns of the developed and validated salivary GC biomarkers, miR-140-5p and miR-301a-3p, were observed in the biofluids of GC patients, supporting the diagnostic relevance of these cargo molecules. Notably, saliva emerged as the most promising biofluid for GC diagnosis, achieving superior receiver-operating characteristic curve values compared with plasma and tissue. This study highlights the role of BEST and MBA in advancing single-EVP analysis and elucidating EVP trafficking, paving the way for future diagnostic applications of EVP cargo.

Antibody-mediated rejection is a leading cause of allograft failure and mortality in pediatric solid organ transplant recipients. Current apheresis systems require large blood volumes and are primarily designed for adults, making them unsuitable for children and small animals. These systems often indiscriminately remove both harmful and protective antibodies, increasing the risk of complications such as life-threatening infections. To address this critical need, we developed acoustofluidic-based system for targeted antibody removal in transplantation (A-START). A-START is engineered to handle small blood volumes and has demonstrated efficacy in preclinical small animal trials. In a sensitized rodent skin transplantation model, A-START only demands 240 microliters of blood, selectively removing donor-specific alloantibodies (DSAs) while preserving protective antibodies, such as tetanus antibodies. A-START retains ~95% of beneficial antibodies and achieves a 60% improvement in DSA removal compared to conventional methods. These findings highlight the transformative potential of A-START as a promising, reliable, scalable solution for improving outcomes in pediatric transplantation and treating antibody-mediated diseases.

Mechanoimmunology explores how mechanical forces orchestrate immune responses, offering insights into immune cell functions and the mechanisms underlying mechanotransduction. A critical challenge in this field is the absence of reliable platforms that apply precise, consistent mechanical stimuli to individual cells while enabling reproducible immune activation. Here, we present a nanoscale acoustic oscillator for mechanoimmunology applications: NAOMI. NAOMI features micropatterned pillars that support uniform cell monolayer formation with an integrated acoustic transducer that delivers highly controlled 3D nanoscale oscillations (±1-nanometer deviation) for up to 72 hours. Unlike conventional passive platforms relying on static stiffness or surface topography, NAOMI enables dynamic, programmable stimulation with high precision and reproducibility. Validation studies demonstrate that NAOMI notably enhances mechanical stress intensity and cell displacement, driving robust M1 polarization in macrophages. NAOMI provides a practical and versatile platform for studying mechanoimmunology, offering high precision, stability, and tunability. Its capabilities also position it well to support future research and drive innovative discoveries in the field.

Extracellular vesicles (EVs) play a crucial role in intercellular communication, signaling pathways, and disease pathogenesis by transporting biomolecules such as DNA, RNA, proteins, and lipids derived from their cells of origin, and they have demonstrated substantial potential in clinical applications. Their clinical significance underscores the need for sensitive methods to fully harness their diagnostic potential. In this comprehensive review, we explore EV heterogeneity related to biogenesis, structure, content, origin, sample type, and function roles; the use of EVs as disease biomarkers; and the evolving landscape of EV measurement for clinical diagnostics, highlighting the progression from bulk measurement to single vesicle analysis. This review covers emerging technologies such as single-particle tracking microscopy, single-vesicle RNA sequencing, and various nanopore-, nanoplasmonic-, immuno-digital droplet-, microfluidic-, and nanomaterial-based techniques. Unlike traditional bulk analysis methods, these methods contribute uniquely to EV characterization. Techniques like droplet-based single EV-counting enzyme-linked immunosorbent assays (ELISA), proximity-dependent barcoding assays, and surface-enhanced Raman spectroscopy further enhance our ability to precisely identify biomarkers, detect diseases earlier, and significantly improve clinical outcomes. These innovations provide access to intricate molecular details that expand our understanding of EV composition, with profound diagnostic implications. This review also examines key research challenges in the field, including the complexities of sample analysis, technique sensitivity and specificity, the level of detail provided by analytical methods, and practical applications, and we identify directions for future research. This review underscores the value of advanced EV analysis methods, which contribute to deep insights into EV-mediated pathological diversity and enhanced clinical diagnostics.

Mechanical forces constantly stimulate cellular functions and influence their response behaviors. Similar to how an orchestra's music synchronizes an audience, acoustic technologies have emerged as precise, contact-free tools to study cellular responses. These platforms generate forces at appropriate length and frequency scales, enabling precise interactions with cells. Recent advancements highlight their potential for regulating cellular functions, revealing both therapeutic promise and the need for further biochemical exploration. This review summarizes the progress in using acoustic technologies to orchestrate cellular functions in vitro through mechanical stimulation. We first introduce the main categories of acoustic platforms and their working principles in cellular research. Subsequently, we explore the fundamental mechanisms linking acoustics to specific cellular interactions. We then review recent applications of these technologies in precisely modulating cellular functions for therapeutic purposes. Last, we discuss strategies to enhance their performance and efficacy, along with their potential integration with other biomedical tools.

, the field of micro/nanorobots has evolved from science fiction to reality, with significant advancements in biomedical and environmental applications. Despite the rapid progress, the deployment of functional micro/nanorobots remains limited. This review of the technology roadmap identifies key challenges hindering their widespread use, focusing on propulsion mechanisms, fundamental theoretical aspects, collective behavior, material design, and embodied intelligence. We explore the current state of micro/nanorobot technology, with an emphasis on applications in biomedicine, environmental remediation, analytical sensing, and other industrial technological aspects. Additionally, we analyze issues related to scaling up production, commercialization, and regulatory frameworks that are crucial for transitioning from research to practical applications. We also emphasize the need for interdisciplinary collaboration to address both technical and nontechnical challenges, such as sustainability, ethics, and business considerations. Finally, we propose a roadmap for future research to accelerate the development of micro/nanorobots, positioning them as essential tools for addressing grand challenges and enhancing the quality of life.

Arthropods have intricate compound eyes and optic neuropils, exhibiting exceptional visual capabilities. Combining the strengths of digital imaging with the features of natural arthropod visual systems offers a promising approach to harness wide-angle vision and depth perception while addressing limitations like low resolving power. Here, we present an artificial intelligence-assisted biomimetic system modeled after arthropod vision. We developed a biomimetic compound eye camera with an effective pixel number of 4.3 megapixels capable of producing full-color panoramic images with a viewing angle of 165° and resolving power of 40 micrometers. Using rich visual information, our system achieves high-fidelity image reconstruction, precise 3D position prediction, high-accuracy classification, and pattern recognition through a multistage neural network. Moreover, our compact biomimetic visual system can simultaneously track the 3D motion of multiple miniature targets independently. The proof-of-concept biomimetic arthropod visual system offers a computational panoramic imaging solution, advancing applications in industry, medicine, and robotics.

Granular hydrogel scaffolds composed of many discrete hydrogel microparticles (HMPs) have demonstrated significant advantages over bulk hydrogels, including injectability and the flexibility to incorporate diverse chemistries, physical properties, and bioactive payloads. Herein, we demonstrate the ability to tune HMP properties through varying the length of poly(ethylene glycol) (PEG) arms and stereocomplexed poly(lactic acid) (SC PLA) cross-links within PEG-based HMPs to further understand the networks' structure-property relationships and utility in a model prodrug delivery system. DSC and WAXS revealed that hydrogels with shorter PEG arms were able to form stereocomplex domains to a greater extent than longer PEG arms. Additionally, as the SC PLA length increased, the HMPs were more thermally and mechanically stable. HMPs were also loaded with model prodrug, doxorubicin, to characterize compositional variations' effects on release profiles. These studies suggest that variations in the cross-linker concentration influence the crystallinity of each HMP formulation, allowing for tunable drug loading and release.

The rotation of objects and corresponding dynamic systems plays a critical role in applications ranging from microscale droplet-based biochemical assays to nanoscale fluid transport and targeted drug delivery. However, directly observing and controlling these rotational phenomena across these different scales remains a challenge. Here, we introduce an acoustofluidic spinning control method that dynamically guides particles into three-dimensional, periodic spatial patterns within a droplet. Using surface acoustic waves, we induce internal streaming that generates centrifugal forces counteracted by surface tension, leading to the formation of rotating Stokes waves along the droplet's equator. We show that fluid motion inside the droplet couples with these rotating waves, giving rise to a controllable superimposed helical particle orbit. These findings provide a platform for controlled rotational flows with potential applications in droplet-based microfluidics, biochemical processing, and tunable particle transport in lab-on-a-chip systems.

Contactless manipulation of samples, particularly the ability to dynamically handle multiple fragile specimens while maintaining their integrity and viability, is crucial for various applications in biology, medicine, engineering, and physics. While hydrodynamic tweezers have emerged as a promising approach for gentle, label-free manipulation of a wide range of sample types and sizes, they typically have limited flexibility in terms of dynamic control, making it challenging to realize high-resolution and programmable manipulation of multiple samples. Here, we introduce the Streaming-based Tweezers for Routing, Engineering, And Manipulation of multiparticles (STREAM) with sub-wavelength resolution. The platform employs an array of piezoelectric plates arranged in a space-reciprocal pattern to generate acoustic streaming, creating localized trapping points. The mechanism of particle trapping and the improvement of routing resolution via multiunit activation were investigated. Subsequently, a convolutional neural network (CNN) with arbitrary voltage combination as the input and predicted trapping position as the output was integrated into the system. The CNN calibration is vital to the system as it enhances the platform's performance, enabling precise control of the trapping positions beyond traditional physical unit size limitations. We demonstrated the STREAM platform's capabilities through single particle routing with sub-wavelength precision, simultaneous manipulation of multiple particles, and on-demand assembly of samples. The STREAM platform opens new possibilities for applications requiring precise and dynamic control of particles and samples, with the potential to advance fields including biology, chemistry, and materials science.

Droplet manipulation technologies play a critical role in many aspects of biochemical research, including in complex reaction assays useful for drug delivery, for building artificial cells, and in synthetic biology. While advancements have been made in manipulating liquid droplets, the capability to freely and dynamically manipulate solid objects across aqueous and oil phases remains unexplored. Here, we develop an acoustofluidic frequency-associated microsphere embedding platform, which enables microscale rapid injection of microparticles from a fluorinated oil into aqueous droplets. By observing different embedding mechanisms at low and high acoustic frequencies, we establish a theoretical model and practical principles for cross-phase manipulations. The proposed system not only enables multi-phase manipulation but also provides contactless control of specific microparticles within various distinctive phases. We demonstrate the acoustic-driven embedding and subsequent on-demand disassembly of hydrogel microspheres. This system indicates potential for reagent delivery and molecule capture applications. It enhances existing droplet manipulation technologies by enabling both multi-phase and cross-phase operations, paving the way for solid-liquid interaction studies in artificial cell research. The capability for intricate multi-phase loading, transport, and reactions offers promising implications for various fields, including in-droplet biochemical assays, drug delivery, and synthetic biology. Cross-phase manipulation holds potential for applications in synthetic biology and drug delivery. Here, authors present an acoustofluidic platform that enables rapid embedding of microparticles from an oil phase into aqueous droplets, offering an effective tool for studying cellular multiphase interactions and related phenomena.

Recent years have witnessed the surge of topological wave phenomena as a versatile platform to engineer exotic wave energy transport which is robust to defects and disorders. Most demonstrations in acoustics remain in a single phase of matter such as solid or air. Here, we introduce the realization of valley phononic crystals for surface acoustic waves and their interaction with fluids in an acoustofluidic setup. It is shown that the interplay between megahertz elastic waves and hydrodynamics where two phases of materials are involved offers rich physics and new engineering potentials of topological matter. By electroplating hexagonal copper pillars on a lithium niobate substrate and adding a liquid layer on top of it, the excited elastic valley spin is transferred at the interface of solid-fluid domains. The interactions lead to valley streaming vortices in the fluid domain that support backward-immune particle transport. In addition, it is found that pressure wells are formed around the small pillars, which enable the concentration of DNA molecules in the nm size range. The studies may open new avenues for applying topological acoustic waves in particle manipulation and life sciences.

The complex interaction of spin, valley and lattice degrees of freedom allows natural materials to create exotic topological phenomena. The interplay between topological wave materials and hydrodynamics could offer promising opportunities for visualizing topological physics and manipulating bioparticle unconventionally. Here we present topological acoustofluidic chips to illustrate the complex interaction between elastic valley spin and nonlinear fluid dynamics. We created valley streaming vortices and chiral swirling patterns for backward-immune particle transport. Using tracer particles, we observed arrays of clockwise and anticlockwise valley vortices due to an increase in elastic spin density. Moreover, we discovered exotic topological pressure wells in fluids, creating nanoscale trapping fields for manipulating DNA molecules. We also found a 93.2% modulation in the bandwidth of edge states, dependent on the orientation of the substrate's crystallographic structure. Our study sets the stage for uncovering topological acoustofluidic phenomena and visualizing elastic valley spin, revealing the potential for topological-material applications in life sciences.

Inspired by counterintuitive water “swelling” ability of the hydrophobic moss of the genus Sphagnum (Peat moss), we prepared a hydrophobic pseudo-hydrogel (HPH), composed of a pure hydrophobic silicone elastomer with a tailored porous structure. In contrast to conventional hydrogels, HPH achieves absorption-induced volume expansion through surface tension induced elastocapillarity, presenting an unexpected absorption-induced volume expansion capability in hydrophobic matrices. We adopt a theoretical framework elucidating the interplay of surface tension induced elastocapillarity, providing insights into the absorption-induced volume expansion behavior. By systematically programming the pore structure, we demonstrate tunable, anisotropic, and programmable absorption-induced expansion. This leads to dedicated self-shaping transformations. Incorporating magnetic particles, we engineer HPH-based soft robots capable of swimming, rolling, and walking. This study demonstrates a unusual approach to achieve water-responsive behavior in hydrophobic materials, expanding the possibilities for programmable shape-morphing in soft materials and soft robotic applications. Shape-changing materials have potential in a range of applications, but these transformations can be challenging to control. Here, the authors report the hydrophobic pseudo-hydrogel, which utilizes absorption-induced expansion via elastocapillarity to enable versatile soft robotic applications.

Precise and rapid disease detection is critical for controlling infectious diseases like COVID-19. Current technologies struggle to simultaneously identify viral RNAs and host immune antibodies due to limited integration of sample preparation and detection. Here, we present acoustofluidic integrated molecular diagnostics (AIMDx) on a chip, a platform enabling high-speed, sensitive detection of viral immunoglobulins [immunoglobulin A (IgA), IgG, and IgM] and nucleic acids. AIMDx uses acoustic vortexes and Gor'kov potential wells at a 1/10,000 subwavelength scale for concurrent isolation of viruses and antibodies while excluding cells, bacteria, and large (>200 nanometers) vesicles from saliva samples. The chip facilitates on-chip viral RNA enrichment, lysis in 2 minutes, and detection via transcription loop-mediated isothermal amplification, alongside electrochemical sensing of antibodies, including mucin-masked IgA. AIMDx achieved nearly 100% recovery of viruses and antibodies, a 32-fold RNA detection improvement, and an immunity marker sensitivity of 15.6 picograms per milliliter. This breakthrough provides a transformative tool for multiplex diagnostics, enhancing early infectious disease detection.

Abstract Gastric cancer (GC) remains a formidable global health challenge, with late-stage diagnosis and high recurrence rates resulting in poor patient outcomes. This study explores the potential of advanced technologies, namely Bessel Beam Excitation Separation Technology (BEST) and multiparametric biochip assay (MBA), to track single extracellular vesicle and particle (EVP) cargo from organs of pathology into biofluids such as plasma and saliva. Using GC as a study model, we conducted high throughput, multiparametric analyses of EVPs derived from plasma, saliva, and tissue samples. Our findings demonstrate the feasibility of these techniques in isolating and characterizing EVPs, revealing consistent EVP morphology and size across biofluids. Furthermore, differential expression patterns of the developed and validated salivary GC biomarkers, miR-140-5p and miR-301a-3p, were observed in GC patient biofluids, supporting the diagnostic relevance of EVP cargo. Notably, saliva emerged as the most promising biofluid for GC diagnosis, achieving superior Receiver Operating Characteristic (ROC) curve values compared to plasma and tissue. This study highlights the role of BEST and MBA in advancing single-EVP analysis and elucidating EVP trafficking, paving the way for future diagnostic applications of EVP cargo.

Optical filtering is an indispensable part of fluorescence microscopy for selectively highlighting molecules labeled with a specific fluorophore and suppressing background noise. However, the utilization of optical filtering sets increases the complexity, size, and cost of microscopic systems, making them less suitable for multifluorescence channel, high-speed imaging. Here, we present filter-free fluorescence microscopic imaging enabled with deep learning-based digital spectral filtering. This approach allows for automatic fluorescence channel selection after image acquisition and accurate prediction of fluorescence by computing color changes due to spectral shifts with the presence of excitation scattering. Fluorescence prediction for cells and tissues labeled with various fluorophores was demonstrated under different magnification powers. The technique offers accurate identification of labeling with robust sensitivity and specificity, achieving consistent results with the reference standard. Beyond fluorescence microscopy, the deep learning-enabled spectral filtering strategy has the potential to drive the development of other biomedical applications, including cytometry and endoscopy.

Small extracellular vesicles (sEVs) have emerged as powerful vectors for liquid biopsy, offering a noninvasive window into the dynamic physiological and pathological states of the body. However, to fully leverage the clinical potential of sEV biomarkers, it is imperative to develop robust and efficient technologies for their isolation and analysis. In this study, we introduce a novel sharp-edge acoustofluidic platform designed for rapid and effective sample preparation, coupled with sensitive detection of specific sEV populations based on their surface markers. Our approach utilizes acoustically activated sharp-edge microstructures to concentrate bead-bound sEVs within the microfluidic device, facilitating immediate visualization by fluorescence microscopy. As a proof of principle, we demonstrate the capability of this portable acoustofluidic chip to selectively isolate and detect epidermal growth factor receptor (EGFR)-expressing vesicles, achieving nearly a 6-fold signal enhancement in EGFR-positive sEVs compared to EGFR-negative populations from sample volumes as small as 50 μl. This advancement not only underscores the potential of our platform for high-sensitivity biomarker detection but also paves the way for its application in isolating organ-specific sEVs. Such capability could be transformative for real-time monitoring of organ function and the simultaneous detection of multiple sEV markers, thereby broadening the scope of diagnostic precision and therapeutic decision-making in clinical practice.

Mechano-chemo cancer treatment is an emerging therapeutic strategy that enhances chemotherapy efficacy by combining chemical agents with mechanical forces to improve drug uptake and overcome resistance. However, current approaches for delivering mechanical forces, including magnetic stress, hydrodynamic shear, and ultrasonic cavitation, suffer from limited tunability, poor spatial precision, and off-target effects, restricting their clinical potential. Here, we introduce ChemoTAP (chemotherapy-enhanced tunable acoustofluidic permeabilization), an acoustofluidic system that utilizes standing surface acoustic waves (SAWs) to achieve highly localized, tunable mechanical stimulation, enhancing tumor cell permeability and improving chemotherapeutic efficiency. By fine-tuning SAW parameters, ChemoTAP transiently modulates membrane permeability by activating mechanosensitive ion channels, leading to cytoskeletal remodeling and a 2.73-fold increase in intracellular calcium ion flux in HeLa cells. This SAW-induced mechanotransduction response synergistically enhances the cytotoxic effects of cisplatin, increasing tumor cell apoptosis by 1.78-fold through mitochondrial membrane depolarization, reactive oxygen species generation, and endoplasmic reticulum stress pathways. Unlike conventional ultrasound-based cavitation methods, ChemoTAP enables precise, non-invasive mechanical stimulation without requiring microbubbles, offering a controllable and scalable alternative for mechano-chemo cancer treatment. ChemoTAP establishes a foundation for further studies in mechanotherapy treatment pathways and promotes the broader integration of acoustics in oncology.

INTRODUCTION: MicroRNAs (miRNAs), packaged within extracellular vesicles (EVs), have been used to interrogate the pathogenesis of preeclampsia and to identify its biomarkers. We have previously shown that miRNA species were differentially expressed in small plasma EVs from women with preeclampsia vs healthy controls. We sought to assess the use of rapid technologies for isolation of plasma and urine EVs from parturients with preeclampsia and determine differences in the expression of selected EV miRNA species. METHODS: We collected blood and urine samples before delivery from parturients with severe preeclampsia vs healthy controls and used size exclusion chromatography (SEC) as an acceptable standard for rapid isolation of plasma EVs. We also isolated urine and plasma EVs using ExoDisc, a rapid nanofiltration technology for EV isolation. All samples were examined using a nanoparticle tracking analyzer, immunoblotting, and RT-qPCR for selected miRNA levels. RESULTS: Whereas the concentration of EVs was higher in the urine from preeclampsia compared to controls, we observed the opposite change in plasma EVs, with no difference in EV size. Comparing the two patient groups for miRNA levels in EVs isolated by ExoDisc or SEC, we found that EV miR-93-5p was upregulated in the plasma and urine of parturients with preeclampsia vs healthy controls. Notably, miR-31-5p was upregulated in SEC- or ExoDisc-isolated plasma EVs, and miR-92-3p was upregulated in or ExoDisc-isolated plasma or urine EVs of parturients with preeclampsia. DISCUSSION: Technologies for rapid analysis of plasma and urine EVs and their miRNA cargo provide complementary information that might be useful for deciphering pathways leading to preeclampsia and biomarkers for this disease.

Controlled intracellular delivery of biomolecular cargo is critical for developing targeted therapeutics and cell reprogramming. Conventional delivery approaches (e.g., endocytosis of nano-vectors, microinjection, and electroporation) usually require time-consuming uptake processes, labor-intensive operations, and/or costly specialized equipment. Here, we present an acoustofluidics-based intracellular delivery approach capable of effectively delivering various functional nanomaterials to multiple cell types (e.g., adherent and suspension cancer cells). By tuning the standing acoustic waves in a glass capillary, our approach can push cells in flow to the capillary wall and enhance membrane permeability by increasing membrane stress to deform cells via acoustic radiation forces. Moreover, by coating the capillary with cargo-encapsulated nanoparticles, our approach can achieve controllable cell-nanoparticle contact and facilitate nanomaterial delivery beyond Brownian movement. Based on these mechanisms, we have successfully delivered nanoparticles loaded with small molecules or protein-based cargo to U937 and HeLa cells. Our results demonstrate enhanced delivery efficiency compared to attempts made without the use of acoustofluidics. Moreover, compared to conventional sonoporation methods, our approach does not require special contrast agents with microbubbles. This acoustofluidics-based approach creates exciting opportunities to achieve controllable intracellular delivery of various biomolecular cargoes to diverse cell types for potential therapeutic applications and biophysical studies.

Abstract Background This study introduces the A utomated H igh‐purity E xosome isolation‐based AD d iagnostics system (AHEADx) . By analyzing and understanding the molecular cargo (proteins and miRNAs) carried by circulating exosomes, researchers found brain‐derived exosome (BDE) levels of P‐S396‐tau, P‐T181‐tau, and Aβ1‐42 are elevated up to 10 years prior to clinical symptoms. Currently, there is no available technology capable of simultaneously isolating and screening exosomal biomarkers for efficient and personalized precision medicine giving early AD diagnosis. Method This NIH funded study will develop and validate AHEADx via integrated acoustofluidics (i.e., the fusion of acoustics and microfluidics) and photonic PCR on‐chip technologies that are capable of fully automated, rapid, precise exosome isolation and accurate analysis for AD diagnostics. AHEADx consists of two units: a rapid (<1 min) acoustofluidic separation unit for exosome isolation from biofluids with high yield and purity (both >90%), and a rapid (<6 min) photonic PCR unit achieving detection limits of ∼1 copy/µL for nucleic acids and ∼5 copies/µL for proteins. Compared with state‐of‐the‐art exosome isolation and analysis technologies, AHEADx system has the following advantages: • Automated and fast operation in a point‐of‐care, handheld system • High‐purity (>90%), high‐quality exosome isolation for accurate biomarker detection • High‐sensitivity (∼1 copy/µL for nucleic acids and ∼5 copies/µL for proteins) detection of a comprehensive (∼20) panel of AD biomarkers Result To validate the potential for clinical use, we will test plasma samples from 100 AD patients and 100 healthy individuals all with known amyloid, phospho tau and total tau biomarker status measured in their CSF. The Duke University Neurology Biobank and the Duke University and University of North Carolina Alzheimer’s Disease Research Center (Duke/UNC ADRC) will provide samples. Conclusion We predict the AHEADx platform will be capable of the simultaneous isolation and analysis of exosome‐derived biomarkers for early AD neuropathological diagnosis. The AHEADx platform’s ability to accurately detect AD biomarkers in the preclinical stages as a point of care handheld instrument could revolutionize diagnosis of AD pathology in the office setting, enhance understanding of AD progression, and significantly impact research into effective treatments.

Abstract Laboratory automation technologies have revolutionized biomedical research. However, the availability of automation solutions at the single-cell level remains scarce, primarily owing to the inherent challenges of handling cells with such small dimensions in a precise, biocompatible manner. Here, we present a single-cell-level laboratory automation solution that configures various experiments onto standardized, microscale test-tube matrices via our precise ultrasonic liquid sample ejection technology, known as PULSE. PULSE enables the transformation of titer plates into microdroplet arrays by printing nanodrops and single cells acoustically in a programmable, scalable, and biocompatible manner. Unlike pipetting robots, PULSE enables researchers to conduct biological experiments using single cells as anchoring points (e.g., 1 cell vs . 1000 cells per “tube”), achieving higher resolution and potentially more relevant data for modeling and downstream analyses. We demonstrate the ability of PULSE to perform biofabrication, precision gating, and deterministic array barcoding via preallocated droplet-addressable primers. Single cells can be gently printed at a speed range of 5–20 cell⋅s −1 with an accuracy of 90.5–97.7%, which can then adhere to the substrate and grow for up to 72 h while preserving cell integrity. In the deterministic barcoding experiment, 95.6% barcoding accuracy and 2.7% barcode hopping were observed by comparing the phenotypic data with known genotypic data from two types of single cells. Our PULSE platform allows for precise and dynamic analyses by automating experiments at the single-cell level, offering researchers a powerful tool in biomedical research.

Advanced biofabrication techniques can create tissue-like constructs that can be applied for reconstructive surgery or as in vitro three-dimensional (3D) models for disease modeling and drug screening. While various biofabrication techniques have recently been widely reviewed in the literature, acoustics-based technologies still need to be explored. The rapidly increasing number of publications in the past two decades exploring the application of acoustic technologies highlights the tremendous potential of these technologies. In this review, we contend that acoustics-based methods can address many limitations inherent in other biofabrication techniques due to their unique advantages: noncontact manipulation, biocompatibility, deep tissue penetrability, versatility, precision in-scaffold control, high-throughput capabilities, and the ability to assemble multilayered structures. We discuss the mechanisms by which acoustics directly dictate cell assembly across various biostructures and examine how the advent of novel acoustic technologies, along with their integration with traditional methods, offers innovative solutions for enhancing the functionality of organoids. Acoustic technologies are poised to address fundamental challenges in biofabrication and tissue engineering and show promise for advancing the field in the coming years.

Ring resonator (RR) devices are closed-loop waveguides where waves circulate only at the resonant frequencies. They have been used in sensor technology and optical tweezers, but controlling micron-scale particles with optical RR tweezers is challenging due to insufficient force, short working distances, and photodamage. To overcome these obstacles, an acoustofluidic RR-based tweezing method is developed to manipulate micro-sized particles that can enhance particle trapping through the resonance interaction of acoustic waves with high Q factor (>3000), more than 20 times greater than traditional acoustic transducers. Particles can be precisely manipulated within the RR by adjusting the signal phase, with trapping amplified by enlarging the connected waveguide. Rapid particle mixing is achieved when particles are placed between the waveguide and RR. The signal path is strengthened by strategically positioning the RR in a two-dimensional plane. Acoustofluidic RR tweezers have immense potential for advancing applications in biosensing, mechanobiology, lab-on-a-chip, and cell-cell communication research.