Lab on a Chip最新文献

Over the past 15 years, smartphones have had a transformative effect on everyday life. These devices also have the potential to transform molecular analysis over the next 15 years. The cameras of a smartphone, and its many additional onboard features, support optical detection and other aspects of engineering an analytical device. This article reviews the development of smartphones as platforms for portable chemical and biological analysis. It is equal parts conceptual overview, technical tutorial, critical summary of the state of the art, and outlook on how to advance smartphones as a tool for analysis. It further discusses the motivations for adopting smartphones as a portable platform, summarizes their enabling features and relevant optical detection methods, then highlights complementary technologies and materials such as 3D printing, microfluidics, optoelectronics, microelectronics, and nanoparticles. The broad scope of research and key advances from the past 7 years are reviewed as a prelude to a perspective on the challenges and opportunities for translating smartphone-based lab-on-a-chip devices from prototypes to authentic applications in health, food and water safety, environmental monitoring, and beyond. The convergence of smartphones with smart assays and smart apps powered by machine learning and artificial intelligence holds immense promise for realizing a future for molecular analysis that is powerful, versatile, democratized, and no longer just the stuff of science fiction.

Biological nanomaterials have unique magnetic and density characteristics that can be employed to isolate them into subpopulations. Extracellular nanovesicles (EVs) are crucial for cellular communication; however, their isolation poses significant challenges due to their diverse sizes and compositions. We present EV-Lev, a microfluidic magnetic levitation technique for high-throughput, selective isolation of small EVs (<200 nm) from human plasma. EV-Lev overcomes the challenges posed by the subtle buoyancy characteristics of EVs, whose small size and varied densities complicate traditional magnetic levitation techniques. It employs antibody-coated polymer beads of varying densities, integrating immuno-affinity and microfluidics to isolate EVs from sub-milliliter plasma volumes efficiently. It facilitates rapid, simultaneous sorting of EV subpopulations based on surface markers, such as CD9, CD63, and CD81, achieving high yield and purity. Subsequent size and morphology analyses confirmed that the isolated EVs maintain their structural integrity. EV-Lev could help uncover the cargo and function of EV subpopulations associated with multiple diseases including cancer, infectious diseases and help to discover potential biomarkers in small volume samples, while offering a portable, cost-effective, and straightforward assay scheme.

Soft actuators have developed over the last decade for diverse applications including industrial machines and biomedical devices. Integration of chemical sensors with soft actuators would be beneficial in analyzing chemical and environmental conditions, but there have been limited devices to achieve such sensing capabilities. In this work, we developed a thin-film soft actuator integrated with a paper-based chemical sensor, termed a microfluidic paper-based analytical soft actuator (μPAC). μPAC consists of (1) a silicone thin film with a 3D-printed pneumatic chamber and (2) a cellulose paper. This cellulose paper offers dual functions: the strain-limiting layer of a soft actuator and the substrate for the chemical sensor for a paper-based analytical device (μPAD). We characterized the design parameters of the actuators-namely, (1) thickness of silicone thin film, (2) chamber length, and (3) Young's modulus of silicone thin film-to evaluate the actuation performance. These characterizations suggested that the cellulose paper served as a suitable self-straining layer of the actuator, making μPAC a chemical sensor that can actuate simultaneously. Highlighting the unique capability of μPAC, we demonstrated the local detection of pH on the curved target surface. Overall, this research demonstrated the rapid fabrication of actuating chemical sensors with a unique design by combining soft actuators and μPAD, enabling chemical sensing on various surface topologies by dynamically making conformal contact.

A longstanding challenge in microfluidics has been the efficient delivery of fluids from macro-scale pumping systems into microfluidic devices, known as the "world-to-chip" problem. Thus far, the entire industry has accepted the use of imperfect, rigid tubing and connectors as the ecosystem within which to operate, which, while functional, are often cumbersome, labor-intensive, prone to errors, and ill-suited for high-throughput experimentation. In this paper, we introduce TapeTech microfluidics, a flexible and scalable solution designed to address the persistent "world-to-chip" problem in microfluidics, particularly in organ-on-a-chip (OoC) applications. TapeTech offers a streamlined alternative, utilizing adhesive tape and thin-film polymers to create adaptable, integrated multi-channel ribbon connectors that simplify fluidic integration with pumps and reservoirs. Key features of TapeTech include reduced pressure surges, easy priming, rapid setup, easy multiplexing, and broad compatibility with existing devices and components, which are essential for maintaining stable fluid dynamics and protecting sensitive cell cultures. Furthermore, TapeTech is designed to flex around the lids of Petri dishes, enhancing sterility and transportability by enabling easy transfer between incubators, biosafety cabinets (BSCs), and microscopes. The rapid design-to-prototype iteration enabled by TapeTech allows users to quickly develop connectors for a wide range of microfluidic devices. Importantly, we showcase the utility of TapeTech in OoC cultures requiring fluid flow. We also highlight other utilities, such as real-time microscopy and a well-plate medium exchanger. The accessibility of this technology should enable more laboratories to simplify design and setup of microfluidic experiments, and increase technology adoption.

Antibody discovery can benefit from techniques to screen antibody-secreting cells (ASCs) at scale for the binding and functionality of a diverse set of secreted antibodies. Previously, we demonstrated the use of cavity-containing hydrogel microparticles (nanovials) coated with a single affinity agent, biotin, to capture and identify ASCs secreting antibodies against a recombinant antigen bound to the nanovial through biotin-streptavidin linkages. However, rapidly secreted antibodies from unbound cells or cells in adjacent nanovials can cause crosstalk leading to background signal. Earlier efforts address this by localizing capture sites to the nanovial cavity, emulsifying nanovials, or short secretion times to limit secreted antibodies from binding to neighboring nanovials. Here, we demonstrate a method to functionalize nanovials with moieties that impart orthogonal reactivity, enabling conjugation of cell capture antibodies and antigens at different times. We show that by using a strained alkyne moiety to attach cell-capture antibodies via click chemistry to nanovials, we can capture cells and subsequently quantify secretions via biotin-streptavidin linkages. By delaying the loading of antigens onto the nanovials until after cell capture, we were able to ensure high purity (>95%) isolation of hybridoma secreting an antigen-specific antibody in a background of other hybridoma. This approach allows tight temporal control of the secretion measurement, which is independent of the cell loading time and requires less convective transfer steps. Click chemistry-based coupling further improved cell loading into nanovials by 58% compared to biotin-streptavidin-biotin coupling and caused no reduction in cell viability. We demonstrate an implementation of this system to improve antigen-specific hybridoma screening, yielding an 8-fold improvement in hybridoma enrichment while maintaining similar workflow complexity. Hybridomas on nanovials sorted into well plates regrew into colonies following sorting using standard fluorescence-activated cell sorting and maintained secretion of antigen-specific antibodies with high purity (∼90%), as validated via standard enzyme-linked immunosorbent assays. This lab-on-a-particle approach can be applied more generally to decouple cell loading, treatment, or activation, from secretion measurements for single-cell functional assays.

Timely identification of infectious pathogens is crucial for the accurate diagnosis, management, and treatment of syndromic respiratory diseases. Nevertheless, the implementation of rapid, precise, and automated point-of-care testing (POCT) remains a significant challenge. This study introduces an advanced digital microfluidic (DMF) POCT testing system designed for the rapid molecular syndromic testing of multiple respiratory pathogens from a single untreated sample. The system seamlessly integrates magnetic beads-based nucleic acid extraction, PCR amplification, and real-time fluorescence analysis in an automatic run, facilitating sample-to-answer detection within 80 min. It accommodates various sample types, including nasopharyngeal swabs, oropharyngeal swabs, bronchoalveolar lavage fluid, and sputum. A facile sample loading method has been developed to reduce hands-on time to less than 1 min. The system exhibits high sensitivity (200-628 copies per mL) for 15 pathogens and has the capacity for up to 32 multiplexed tests per run. Validation with 255 clinical samples confirms its high sensitivity and specificity. The DMF-based system significantly reduces manual labour, enhances rapid POCT for respiratory infections, and, with optimized manufacturing processes, lowers costs for large-scale production. The system can be applied and improve clinical management near the patients as well as in resource-limited settings.

Bone related disorders are highly prevalent, and many of these pathologies still do not have curative and definitive treatment methods. This is due to a complex interplay of multiple factors, such as the crosstalk between different tissues and cellular components, all of which are affected by microenvironmental factors. Moreover, these bone pathologies are specific, and current treatment results vary from patient to patient owing to their intrinsic biological variability. Current approaches in drug development to deliver new drug candidates against common bone disorders, such as standard two-dimensional (2D) cell culture and animal-based studies, are now being replaced by more relevant diseases modelling, such as three-dimension (3D) cell culture and primary cells under human-focused microphysiological systems (MPS) that can resemble human physiology by mimicking 3D tissue organization and cell microenvironmental cues. In this review, various technological advancements for in vitro bone modeling are discussed, highlighting the progress in biomaterials used as extracellular matrices, stem cell biology, and primary cell culture techniques. With emphasis on examples of modeling healthy and disease-associated bone tissues, this tutorial review aims to survey current approaches of up-to-date bone-on-chips through MPS technology, with special emphasis on the scaffold and chip capabilities for mimicking the bone extracellular matrix as this is the key environment generated for cell crosstalk and interaction. The relevant bone models are studied with critical analysis of the methods employed, aiming to serve as a tool for designing new and translational approaches. Additionally, the features reported in these state-of-the-art studies will be useful for modeling bone pathophysiology, guiding future improvements in personalized bone models that can accelerate drug discovery and clinical translation.

Quantifying infectious virus is essential for vaccine development, clinical diagnostics, and infectious disease research, but current assays are constrained by long turnaround times, high costs, and laborious procedures. To address these limitations, we present a digital focus assay employing an array of independent nanoliter cell cultures. The microfluidic platform allows cells in each nanowell to be inoculated with virus, followed by oil discretization to prevent cross-contamination. After incubation, infected cells are visualized through immunofluorescence staining, and a binary map of wells positive for viral antigen is generated by automated image analysis, allowing infectious viral titer to be calculated by statistical analysis. The platform requires significantly smaller sample and reagent volumes than conventional focus assays while enhancing assay automation and endpoint time flexibility. The technology is applied to the quantification of infectious influenza A using both model virus and clinical specimens, demonstrating the digital platform as an accurate, rapid, cost-effective, and convenient tool for viral load quantification with broad utility in clinical, pharmaceutical, and research applications.

Recent advancements in single-cell analysis have underscored the need for precise isolation and manipulation of individual cells. Traditional techniques for single-cell manipulation are often limited by the number of cells that can be parallel trapped and processed and usually require complex devices or instruments to operate. Here, we introduce an acoustic microfluidic platform that efficiently traps and selectively releases individual cells using spherical air cavities embedded in a polydimethylsiloxane (PDMS) substrate for large scale manipulation. Our device utilizes the principle of acoustic impedance mismatches to generate near-field acoustic potential gradients that create trapping sites for single cells. These single cell traps can be selectively disabled by illuminating a near-infrared laser pulse, allowing targeted release of trapped cells. This method ensures minimal impact on cell viability and proliferation, making it ideal for downstream single-cell analysis. Experimental results demonstrate our platform's capability to trap and release synthetic microparticles and biological cells with high efficiency and biocompatibility. Our device can handle a wide range of cell sizes (8-30 μm) across a large active manipulation area of 1 cm² with 20 000 single-cell traps, providing a versatile and robust platform for single-cell applications. This acoustic microfluidic platform offers a cost-effective and practical method for large scale single-cell trapping and selective releasing with potential applications in genomics, proteomics, and other fields requiring precise single-cell manipulation.

Antimicrobial stewardship plays an essential role in combating the global health threat posed by multidrug-resistant pathogens. Phenotypic antimicrobial susceptibility testing (AST) is the gold standard for analyzing bacterial responses to antimicrobials. However, current AST techniques, which rely on end-point bulk measurements of bacterial growth under antimicrobial treatment in a broth solution, have limitations in resembling the physiological working environment and resolving heterogeneity in response kinetics within the population. In this study, we investigate the responses of uropathogenic bacteria under antimicrobial treatment in individual urine. Our results demonstrate substantial heterogeneity in time–kill kinetics in response to antimicrobials in a host-dependent manner. We also establish a microfluidic gel encapsulation platform for single cell imaging to rapidly resolve heterogeneous subpopulations in response to antimicrobials. The platform captures both bacterial growth and killing within the gel and enables medium exchange to assess the ability of surviving cells to resume growth after antimicrobial removal. Our study lays the foundation for a new generation of precision single cell analysis for personalizing antimicrobial treatment.