Abstract Biological systems dynamically grow high-aspect-ratio architectures from a site, enabling traversal of phase boundaries and functional execution. Emulating this growth strategy in synthetic systems could yield functional microsystems for operation across interfaces. However, engineering such bio-inspired growth to proceed out of plane from a substrate in synthetic colloidal assemblies remains challenging, as it requires overcoming gravitational collapse while maintaining structural coherence during extension. Here, we present a field-driven particle system that achieves gravity-resisting growth of high-aspect-ratio structures via frequency-modulated magnetic and hydrodynamic interactions. This growth is enabled by combining static and oscillating magnetic fields, which guide the assembly of magnetic particles into dynamic structures exhibiting a distinct segmented, seaweed-like morphology. These architectures are reconfigurable, stabilizable, programmably actuatable, and capable of penetrating a perfluorohexane–water interface. When functionalized with enzymes, the growing structures act as micro-transporters, delivering catalytic activity across the interface and triggering detectable reactions in both bulk two-phase and microfluidic chip systems. This work establishes a field-driven assembly-to-function approach that integrates structural growth, phase-boundary penetration, and triggered functionality, enabling active microsystems capable of interfacial transport and functional execution.

Acoustofluidics uses engineered acoustic fields to manipulate cells, vesicles, biofluids and tissues in a contact-free and highly programmable manner across microscale to macroscale biological systems. Originally developed as a physics-driven platform for microscale manipulation, acoustofluidics is increasingly emerging as a clinically relevant technology for diagnostics, therapeutics and physiological monitoring. However, clinical translation remains limited by insufficient component standardization, limited system integration beyond the proof-of-concept stage and the lack of established regulatory pathways. This Review addresses this gap by offering a structured discussion of translational progress in acoustofluidics, organized around five key healthcare and medical workflows: biomarker isolation and manipulation, point-of-care diagnostics, in vivo therapeutic applications, wearable and implantable acoustic systems, and end-to-end translation pathways from laboratory prototypes to clinically deployable products. Acoustofluidics is emerging as a versatile platform for clinical diagnostics and therapy, enabling real-time, label-free sensing, manipulation and control of biological and chemical systems across scales. This Review discusses translational progress across key medical workflows, highlights challenges in standardization and integration, and outlines pathways from proof-of-concept devices to clinically deployable systems.

The isolation of small extracellular vesicles (sEVs), viruses and other nanoscale lipid particles from biofluids offers actionable possibilities for advancing disease diagnosis, drug delivery, regenerative medicine, personalized medicine and immunotherapy. Several methods are available to isolate sEVs from biofluids and acoustic techniques provide distinct advantages. Challenges constraining its wider application encompass the absence of adequate procedures for fabrication, implementation and performance validation. These issues impede the development of protocols applicable to nanoscale bioparticles experiencing acoustic isolation effects. Here we present a detailed protocol for acoustic separation of nanoscale bioparticles from biofluids, including plasma and saliva, achieving both high purity and throughput suitable for routine application. This protocol offers a comprehensive, step-by-step guide for the design and fabrication of the acoustic separation device, the establishment of the experimental setup and the isolation of bioparticles. To ensure reliability, rigor and reproducibility, we delineate essential procedures, including acoustic field optimization, channel fabrication and biofluid preparation, subsequently validating the protocol and its performance across different operators. Our protocol further encompasses procedures for data collection and analysis, which are essential for characterizing viruses and sEVs, as well as for evaluating their quality and integrity. This protocol enables researchers to perform high-quality isolation of nanoscale bioparticles, providing access to reliable acoustic separation techniques. Standardizing this technique will pave the way for discoveries in virology and intercellular communication research, with applications in medicine, biology, and materials science. This protocol covers the acoustics-based separation of biological particles such as viruses and single extracellular vesicles from biofluids, including plasma and saliva, with high purity.

Integrated microfluidic biosensors have rapidly evolved into powerful platforms to meet the increasing demand for ultrasensitive and high-throughput quantitative analysis. By seamlessly combining sample handling through microfluidics with real-time detection via 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.

Malignancies of the nervous system remain a critical challenge in oncology, contributing substantially to cancer-related mortality in children and adults. Their aggressive behavior is influenced by adhesion heterogeneity of the tumor cells, as distinct patterns of cell-to-cell and cell-to-matrix interactions shape tumor organization, invasion, and resistance to conventional treatments. Here, we investigate tumor adhesion dynamics in a 3D spherical cavity culture platform that enables the rapid and high-throughput formation of uniform spheroids for parallel morphogenesis, adhesion profiling, and drug screening. The morphogenetic patterns of human neuroblastoma and patient-derived glioblastoma observed in 3D cavity culture closely correlated with tumor aggressiveness, ranging from adhesive layers in benign tumor subtype to compact spheroids in malignant cells and irregular aggregates in highly invasive, cancer stem-like counterparts. These morphologies corresponded to adhesion profiles: aggressive cells displayed elevated N-cadherin with variable integrin, while compact spheroids maintained a balance of both molecules. Blocking N-cadherin with ADH-1 disrupted spheroid integrity, and combining ADH-1 with DOX yielded synergistic cytotoxic effects in malignant phenotypes, highlighting N-cadherin upregulation in tumor cells with higher aggressiveness in 3D culture. Overall, our dynamic 3D spherical cavity culture enables reproducible 3D adhesion phenotyping, providing a simple method for malignancy assessment and supporting adhesion-targeted therapeutic strategies for personalized medicine.

Preterm labor is a serious concern that can lead to preterm birth, posing substantial risks for both the mother and the neonate. Despite approximately 15 million preterm births worldwide each year, there is a lack of sufficient strategies for predicting and preventing preterm labor. Here, we present a non-invasive method for simultaneously detecting exosomal miRNA and protein biomarkers in vaginal discharge, enabling early diagnosis of life-threatening conditions in both the mother and the neonate. Our non-invasive vaginal discharge biopsy using a swab enables the isolation of enriched intact extracellular vesicles through our microfluidic platform called Biologically-intact Exosome Separation Technology (BEST). We observed differential expression of specific miRNAs, including up-regulated hsa-miR-206 and down-regulated hsa-miR-3674, hsa-miR-365a-5p, and hsa-miR-193b-3p, in mothers experiencing preterm labor. We also found significant differences in protein expression in mothers with preterm labor compared to full-term mothers, indicating the involvement of HGS, ATL3, APOH, and GUSB in preterm labor mechanisms. We envision a future in which non-invasive detection of unique miRNA and protein biomarkers in vaginal discharge transforms global healthcare by enabling early detection and effective treatment of preterm labor.

ABSTRACT Severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2) has underscored an urgent need for rapid, accurate, and accessible diagnostic tools to detect infections, facilitate timely quarantine, and inform therapeutic decisions. Fabricating nanostructures on biosensors can enhance biomolecule orientation, minimize steric hindrance, and reduce non‐specific binding, resulting in improved signal‐to‐noise ratios and high sensitivity, making them promising point‐of‐care diagnostic. However, clinical translation remains limited by challenges in scalable and reproducible fabrication and insufficient diagnostic validation. Here, we introduce a nanowell biosensor (NW‐Biosen) fabricated via semiconductor manufacturing, enabling scalable production and yields >1000 electrodes with reproducible properties. NW‐Biosen detects SARS‐CoV‐2 antigens from patient nasal swabs within ∼10 min, with high sensitivity and minimal interference from other coronavirus recombinant proteins, respiratory pathogens, bacteria, and environmental substances, while maintaining consistent reproducibility with different batches. We validated NW‐Biosen through two independent clinical trials involving 249 retrospective and 243 prospective patient samples. In prospective cohort, NW‐Biosen achieved 93.02% sensitivity, 98.73% specificity, and a Cohen's kappa of 0.927, indicating near‐perfect agreement with RT‐PCR and superior sensitivity compared to commercially available colorimetric kits. Thus, NW‐Biosen enables rapid, highly sensitive, reproducible, and cost‐effective at‐home detection, with real‐time data transmission to public health authorities via mobile app integrated with a miniature potentiostat.

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.

Several epidemiological studies have suggested a possible link between Helicobacter pylori (HP) infection and neurodegenerative diseases. However, in the current research, the mechanistic basis of this association remains unclear due to the complexity of multiple effects triggered by HP infection. Here, we expose in vitro cellular models to HP’s cell-free supernatant ( HP CFS) and perform liquid chromatography–mass spectrometry and inhibition assays to identify potential neurodegenerative risk factors associated with HP. We first confirm that HP CFS disrupts the integrity of the gut and brain barriers via vacuolating cytotoxin A (VacA) binding to low-density lipoprotein receptor-related protein-1 (LRP1), reducing ZO-1 tight junction protein levels and transmembrane electrical resistance. In addition, HP CFS activates central innate immune cells, as evidenced by increased expression of inflammatory markers, elevated oxidative stress, the release of neurotoxic factors from microglia and astrocytes. VacA hinders microglial amyloid-beta phagocytosis by blocking LRP1, a key receptor for amyloid-beta clearance. We observe that HP CFS induces neurodegeneration, as indicated by the presence of phosphorylated tau, phosphorylated alpha-synuclein, synaptic impairment, and neuronal damage. Notably, microglia stimulated with HP CFS exacerbate neurodegeneration more than direct HP CFS exposure to neurons. Furthermore, we demonstrate that neuroinflammation and neurodegeneration mediated by HP CFS are mitigated by an LRP1 antagonist or VacA immunodepletion. This study reveals a cellular mechanism of neurotoxic inflammation through multicellular interactions, paving the way for future therapeutic interventions in HP infection-associated neurodegeneration. Graphical Abstract

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. 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 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. 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.

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.

The effects of magnetic fields on quantum entangled biomolecules are detected using a highly sensitive magneto-fluorescence fluctuation microspectroscopy technique. This marks an important advance in experimental quantum life sciences at the single-photon level.

Noninvasive monitoring of Alzheimer's disease (AD) biomarkers is essential for early diagnosis and treatment efficacy. However, noninvasive monitoring of tau protein secretion, a key biomarker of AD, across developmental stages, age‐related variations, and the interaction between apolipoprotein E ( APOE ) and the tau protein axis is not yet accomplished. Here, the label‐free and non‐invasive detection of multiple tau variants dynamics across developmental stages, age‐related variants, and various APOE isogenic genotyes is presented to investigate the APOE –tau axis using human cerebral organoids (hCOs) combined with surface‐enhanced Raman spectroscopy (SERS). Principal component analysis (PCA) of SERS signals successfully identifies four developmental stages of hCOs: embryonic body, neuronal differentiation, maturation, and maintenance phases. Temporal dynamics of age‐related tau protein secretion are observed, reflecting characteristics associated with AD, which are diminished by astrocyte expression. PCA‐based dimensionality reduction of SERS signals further reveals distinct clustering for different APOE isogenic genotypes, with tau protein secretion increasing from APOE2/E2 to APOE4/E4 , providing direct insight into the APOE –tau axis in AD. This study introduces a novel method for the non‐invasive clinical assessments of disease conditions, dynamics, and the relationship between APOE and tau in AD.

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.

Due to a pivotal role in the post-transcriptional regulation of genes implicated in numerous diseases, miRNAs serve as promising disease biomarkers and therapeutic targets. We introduce a new oligonucleotide probe termed miRNA-trigger, which selectively downregulates newly assigned target mRNAs by hijacking specific miRNAs. By engineering the miRNA-trigger to suppress the anti-apoptotic BCL-xL gene, we induce apoptosis selectively in breast cancer cells overexpressing specific miRNAs and further validate its therapeutic efficacy in vivo, by significantly reducing the tumor volume of the xenograft mouse upon its tail-vein injection. This approach establishes a new platform for self-modulating oligonucleotide therapy by redirecting disease-associated miRNAs.

Piezo1 is a mechanosensitive ion channel that has been recently recognized to mitigate Alzheimer’s disease pathological signatures such as beta-amyloid (Aβ) deposition and microglial proinflammation. However, much less is known on how Piezo1 regulates tauopathy, partly due to lack of Alzheimer’s disease (AD) model to elucidate the complex neuroglia interactions. Here, we report how the activation of Piezo1 triggers an anti-inflammatory response and reduces tauopathy using our human Alzheimer’s disease model. We first observed that the activation of Piezo1 suppressed microglial nuclear factor-kB (NFκB), while promoting Aβ clearance and reducing oxidative stress. Piezo1 also reduced astrocytic proinflammation driven by Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3), while promoting Aβ clearance and reducing oxidative stress. We discovered that Piezo1 facilitated tau dephosphorylation and reduced tau hyperphosphorylation by independently enhancing protein phosphatase 2A (PP2A) and inhibiting cyclin-dependent kinase 5 (CDK5) in neurons. Our human AD model demonstrated that Piezo1 served as a crucial mechanosensor in alleviating tauopathy and glial proinflammation, providing a potential therapeutic target in tauopathy-mediated neurodegeneration for regenerative medicine.

ABSTRACT The worldwide prevalence of metabolic dysfunction‐associated steatotic liver disease (MASLD) and alcohol‐associated liver disease (ALD) is rising significantly. However, effective treatment methods for these diseases are lacking, and using exosomes as a promising approach faces significant challenges due to their biological integrity. Here, we report the therapeutic effects of biologically intact exosomes derived from healthy hepatic cells in reducing lipids in cells affected by steatotic liver disease. We first obtained biologically intact exosomes, with a peak size of 30–200 nm and a round shape, as determined by CD63 and CD81 exosome markers, using the biologically intact exosome separation technology (BEST). We induced steatotic liver disease models containing lipid droplets in AML12 cells using oleic acid and ethanol, maintaining approximately 90% cell viability compared to normal hepatocytes. Furthermore, we verified the effectiveness of therapeutic exosomes containing two miRNAs (hsa‐miR‐122‐5p and hsa‐miR‐27a‐3p), which significantly reduced lipid accumulation by up to 92.7% in MASLD and 93.2% reduction in ALD at a dosage of 100 µg/mL of intact exosomes over 72 h. Ultimately, these findings highlight the potential of exosomes as a cell‐free treatment for reversing steatotic liver diseases.

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.