Cancer remains one of the leading global health burdens, demanding swift, precise, and accessible diagnostic responses that can overcome the limitations of diagnostic tests performed in centralized laboratory settings. Point-of-care (POC) molecular diagnostics are emerging as transformative tools, enabling decentralized testing across hospitals, outpatient clinics, oncology practices, and dental practices, thereby accelerating clinical decision-making to improve patient outcomes. This critical review provides a comprehensive perspective on the clinical requirements that shape the design of POC diagnostics, where several case studies based on clinical scenarios serve as representative examples. We detail advancements in POC-compatible sample preparation, nucleic acid and protein biomarker detection, and imaging modalities, highlighting their translational value for real-world oncology applications. In addition, we analyze the global funding landscape, intellectual property, and regulatory frameworks that influence the transition of emerging technologies from academic research to commercial markets. Key enabling technologies, ranging from microfluidics and lab-on-a-chip platforms to nanomaterial-enhanced biosensors and smartphone-integrated devices, are critically evaluated alongside their limitations and challenges. Finally, we project future directions for POC molecular diagnostics, emphasizing interdisciplinary innovation, equitable access, and integration into precision medicine. By synthesizing clinical, technological, and regulatory perspectives, this review underscores the pivotal role of POC molecular diagnostics in shaping the future of cancer detection, therapy selection, therapy effectiveness monitoring, and personalized medicine.

Imaging flow cytometry demands a careful balance between spatial resolution, spectral multiplexing, throughput, and system complexity. Here, we present LFC-plus, a next-generation light-field cytometry platform that enables multiparametric, simultaneous multi-color, and volumetric single-cell analysis. The system integrates model-based image restoration, custom-designed light-field optics, and spectral aperture partitioning, achieving subcellular resolution in all three dimensions, a near-millimeter-scale imaging cross-section, and an analytical imaging throughput of nearly 200 000 cells per second. We validate its performance across diverse biological applications, including chemotherapy response profiling, PEG-mediated cell fusion, and stiffness-based flow migration. These results establish LFC-plus as a robust and scalable platform for high-content volumetric cytometry, with broad implications spanning fundamental biology and translational diagnostics.

Storage options for extracellular vesicles (EVs) remain limited, constraining their clinical and research applications. Conventional low-temperature freezing (-80 °C), although the established standard, requires substantial resources and presents logistical challenges. Here, we report an AridEx (Ambient Retention in Disc of Extracellular vesicles) approach for efficient room-temperature storage and recovery of EVs. AridEx leverages isothermal vitrification principles to preserve EVs, utilizing an electrospun trehalose-dextranol matrix as a xeroprotectant. The xeroprotectant matrix, combined with highly gas-permeable membranes, was integrated into a centrifugal microfluidic device, enabling the rapid desiccation of EV samples under mild vacuum conditions. The dried samples were stored directly on the disc at ambient temperature and subsequently recovered on demand via on-disc rehydration and EV isolation. AridEx achieved preservation efficacy equivalent to that at -80 °C, preserving EV particle counts and surface proteins during ambient storage periods of at least 6 months. In a pilot clinical study, AridEx-preserved plasma retained consistent biomarker signals for accurate cancer diagnosis. This compact and cartridge-based platform is manufacturable and could offer a sustainable alternative to low-temperature sample storage for diagnostic applications.

Whole-organism toxicity assays are essential for evaluating the safety of chemicals, but mammalian models remain costly and low throughput, contributing to a global backlog of untested chemicals. The micro-organism C. elegans has emerged as one of the new approach methodologies (NAMs) modelling toxicological responses across multiple interacting tissues, with advantages of human-relevant biology and cost-effectiveness. However, existing technologies for C. elegans toxicity testing are limited by poor compatibility with long-term chemical exposure, lack of multiplexibility, and inability to dynamically modulate chemical exposure over time. Here, we present a novel microfluidic platform, passive nematode culture chambers (PNCs) integrated into well plates - where micropillar arenas housing crawling or swimming C. elegans have fluid communication with the surrounding medium, enabling consistent nutrient access and long-term viability. These integrated well plates allow multiplexed assays and high-quality imaging due to worm motion being restricted to two-dimensions. We demonstrate the utility of these microfluidic well plates for whole-organism lethality assays. Dose-response studies were conducted with 11 chemicals, revealing median lethal concentrations (LC50s) consistent with values obtained in existing liquid culture and microfluidic systems. The LC50 values from PNC well plates showed strong concordance with median lethal dose data from rats (Spearman correlation, r = 0.827), suggesting that the platform is informative for comparative toxicity ranking. We demonstrate the uniqueness of the PNC well plates by (i) incorporating arenas of varied micropillar spacing and showing that crawling and swimming worms respond differently to heavy metal toxicity due to differences in toxin uptake, (ii) conducting dynamic exposure studies using cadmium chloride (CdCl2), uncovering distinct temporal toxicity patterns and distinguishing between interrupted and chronic exposure insights that would be difficult to obtain using conventional technologies. These findings establish PNC microfluidic well plates as a robust platform for C. elegans-based toxicology. Beyond their immediate application in chemical safety assessment, PNC well plates also enable new experimental directions where minimal intervention, multi-day viability, and scalability are critical.

Immunosenescence dramatically reduces cancer vaccine efficacy in elderly patients, who represent the majority of cancer cases. Despite this clinical reality, the age-related immune decline is rarely considered in preclinical testing. Therefore, novel in vitro models to test cancer vaccine efficacy, considering immunosenescence, are needed. Our novel lymph node paracortex-inspired on-a-chip (LNPoC) platform addresses this gap by recapitulating age-dependent immune responses against cancer vaccines, specifically antigen presentation, antigen-specific T cell activation, and antitumoral responses. Using this platform, we demonstrated that bone marrow-derived antigen-presenting cells (APCs) from young mice (6-7 weeks) displayed significantly enhanced ovalbumin (OVA) peptide presentation compared to APCs from older mice (35-36 weeks). This age-dependent difference translated to significantly greater OVA-specific CD8+ T cell activation and increased cytotoxicity against B16-OVA cancer cells. These age-dependent differences are unique to our LNPoC and undetectable in traditional 2D cultures, confirming that our LNPoC was more effective than 2D cultures at recapitulating immunosenescence-mediated immune responses against cancer vaccines in vitro. The in vivo validation confirms these findings, as young mice demonstrated higher OVA-specific CD8+ T cell responses and smaller tumors than older mice. Our LNPoC is a valuable tool for assessing immunosenescence's impact on cancer vaccines, potentially guiding more effective therapies for older adults.

Conventional electroporation techniques are often limited by low throughput, high reagent consumption, and, critically, the requirement for cell suspension. The latter necessitates enzymatic detachment, subjecting cells to significant physiological stress. To address these challenges, we present a combinatorial screening platform, the programmable digital electroporation system (PDES), which integrates digital microfluidics (DMF) with disposable, flexible interdigitated electrode (IDE) arrays. To enhance system versatility and optical monitoring, we implemented two distinct top-electrode configurations: a flexible polyimide electrode and a large-area transparent quartz electrode. This design enables the fully automated generation of reagent gradients and parallel in situ electroporation of adherent cells under low-voltage conditions. By decoupling fluidic actuation from electrical stimulation, the system facilitates high-throughput parameter optimization while reducing reagent consumption by an order of magnitude. We validated the platform by automatically screening eGFP mRNA transfection conditions, achieving a transfection efficiency of over 90% with cell viability exceeding 95%. Furthermore, we demonstrated the platform's utility in reprogramming human dermal fibroblasts into induced pluripotent stem cells (iPSCs) via circular RNA (circRNA) delivery. Crucially, the PDES performs this process entirely in situ, circumventing the detrimental effects of enzymatic digestion (e.g., trypsinization) inherent to bulk electroporation, the step that often compromises viability and efficiency in sensitive cell types. This work offers a powerful, automated solution for non-viral gene editing, particularly for engineering fragile cells in regenerative medicine and synthetic biology.

Implantable drug delivery systems offer the promise of on-demand, tunable release profiles tailored to individual therapeutic needs. Here, we present a nanofluidic membrane-based electrochemical delivery system that leverages controlled in situ gas generation to achieve electrically modulated molecular transport. The device comprises a monolithically fabricated nanochannel membrane coated with a platinum layer, which enables cathodic water reduction upon application of a −2 VDC potential. This process generates bubbles that transiently increase local pressure, enhancing convective drug transport through the nanochannels. Electrochemical characterization revealed stable gas evolution dynamics with an average actuation current of 2.31 ± 0.36 mA and low power requirements (4.62 ± 0.43 mW), highlighting suitability for energy-constrained implantable settings. In vitro and simulated physiological clearance studies demonstrated reversible, voltage-dependent modulation of drug release across a range of compounds with diverse hydrodynamic radii and charges. Drug release rates ranged from 1 to 10 μg h−1 under electrical actuation—values within therapeutically relevant dosing windows for a wide array of clinical applications. Integration and in vitro validation with a miniaturized Bluetooth-enabled printed circuit board (PCB) controller powered by a 3 V coin cell battery further supports the platform's feasibility for autonomous, wirelessly controlled therapeutic administration. Together, these findings demonstrate a scalable, low-power, and highly adaptable nanofluidic system capable of tunable drug delivery, suitable for integration within implantable closed-loop systems.

Shape-based separation of micro- and nanoparticles has emerged as a powerful yet underdeveloped strategy in microfluidics, offering distinct advantages over conventional size-based methods, particularly for biomedical and functional material applications. Unlike size-based separation, shape-based approaches enable discrimination between particles of identical volume but differing morphology, an essential capability for isolating pathological cells, engineered particles, or anisotropic biological entities whose function is inherently linked to shape. This review provides a comprehensive and critical overview of recent progress in both passive and active microfluidic platforms tailored for shape-selective separation. Passive systems such as deterministic lateral displacement, pinched flow fractionation, inertial, and viscoelastic microfluidics exploit hydrodynamic and flow-structure interactions, while active methods including dielectrophoresis, magnetophoresis, optophoresis, and acoustophoresis utilize external fields to modulate particle trajectories based on geometric anisotropy. For example, recent advancements demonstrate high purities often exceeding 95%, with throughput rates ranging from several microliters to milliliters per minute depending on the device configuration, achieving shape-based cell and particle sorting efficiencies above 90% under optimal conditions. For each technique, we highlight the underlying mechanisms enabling shape sensitivity, key technological advancements, and emerging trends in experimental and computational approaches. We also discuss the challenges in capturing complex particle behaviors such as rotation, alignment, and deformability and emphasize the need for integrated modeling, real-time control, and system-level optimization. Finally, we outline future directions and opportunities for advancing shape-based microfluidic separation toward scalable, high-precision applications in diagnostics, therapeutics, and materials science.

Acoustic microstreaming vortices enable high-throughput cell trapping and sequential delivery of plasmid DNA and Cas9 RNP, boosting transfection up to sevenfold by eliminating cargo competition and/or interaction.

An online, micro free-flow electrophoresis (μFFE) assay was developed for the real-time measurement of insulin in a continuous, competitive immunoassay. Fluorescently labeled insulin and anti-insulin monoclonal antibody were mixed with the sample stream online and flowed through an incubation capillary into the μFFE device. Under an electric field, the bound antibody : insulin-FITC complex was separated from free insulin-FITC. When unlabeled insulin was introduced, labeled and unlabeled insulin competed for binding sites on the antibody, and the separated peaks of the bound complex and free insulin-FITC responded in real-time. Temporal resolution of 30 seconds was achieved following injections of unlabeled insulin, with a limit of detection of 63 nM and a linear response over the tested range of 50 nM to 1 μM insulin. The development of this assay supports the eventual integration of an organ-on-a-chip system capable of measuring direct cellular efflux in tandem with the affinity μFFE detection system. To approximate such an online cellular response, glucose-stimulated insulin secretion was collected offline from human islets and injected into the affinity μFFE instrument. A measurable change in insulin secretion was observed between samples exposed to low and high glucose, demonstrating sufficient limit of detection, linearity, the compatibility of the instrument with a complex biological matrix.