Microfluidic Analysis System Research Articles

Microfluidics for foodborne bacteria analysis: Moving toward multiple technologies integration.

Food security related to bacterial pathogens has seriously threatened human life and caused public health problems. Most of the reported methods are targeted at known major pathogens commonly found in food samples, but to some extent, they have the disadvantage of lacking simplicity, speed, high throughput, and high sensitivity. Microfluidics has become a promising tool for foodborne bacteria analysis and addresses the above limitations. In this perspective, we briefly discussed the ongoing research and development in this field. We outline the major types of microfluidics, the strategies of target biorecognition, and signal amplification technologies in the microfluidic system for the foodborne bacteria analysis. We also proposed the future directions of microfluidics for foodborne bacterial analysis, which aims to integrate multiple technologies toward intelligent analysis with high selectivity and sensitivity for unknown samples, multiple bacterial detection, and simultaneous detection of multiple food pollutants.

Fabrication of Conical Microstructure Array for Stable Droplet Generation Over Wide Flow Rate Range.

Droplet generators with the ability to resist flow fluctuations are of importance for microfluidic chip analysis systems. However, obtaining stably desired-size droplets is still a bugbear since even slight fluctuations can cause polydisperse droplets. In this study, a high-performance droplet generator is achieved with a functional conical array housed in the junction of the channels. The conical microstructures are fabricated through the selective etching of the scratched silicon nitride/silicon (Si3N4/Si) substrate in potassium hydroxide (KOH) etchant, where the combination of lateral and normal material removal contributes to the structure formation. It is found that the key role of the conical microstructures is to regulate the flow rate of the continuous phase, which allows droplet generation to turn to the necking phase and enables droplets to shed more easily. It is also noted that the droplet generator with such a conical array can produce monodisperse droplets in wide-range flow, providing new insights for high-quality device design.

AI-Powered Microfluidics: Shaping the Future of Phenotypic Drug Discovery.

Phenotypic drug discovery (PDD), which involves harnessing biological systems directly to uncover effective drugs, has undergone a resurgence in recent years. The rapid advancement of artificial intelligence (AI) over the past few years presents numerous opportunities for augmenting phenotypic drug screening on microfluidic platforms, leveraging its predictive capabilities, data analysis, efficient data processing, etc. Microfluidics coupled with AI is poised to revolutionize the landscape of phenotypic drug discovery. By integrating advanced microfluidic platforms with AI algorithms, researchers can rapidly screen large libraries of compounds, identify novel drug candidates, and elucidate complex biological pathways with unprecedented speed and efficiency. This review provides an overview of recent advances and challenges in AI-based microfluidics and their applications in drug discovery. We discuss the synergistic combination of microfluidic systems for high-throughput screening and AI-driven analysis for phenotype characterization, drug-target interactions, and predictive modeling. In addition, we highlight the potential of AI-powered microfluidics to achieve an automated drug screening system. Overall, AI-powered microfluidics represents a promising approach to shaping the future of phenotypic drug discovery by enabling rapid, cost-effective, and accurate identification of therapeutically relevant compounds.

Concept, simulation, and fabrication of inverted grating structures for surface plasmon resonance sensors

Abstract. Surface plasmon resonance (SPR) sensors offer the possibility of label-free analysis of biosamples. The long-term stability of standard approaches is limited due to degradation of the grating upon contact with the analyte, and strategies to improve the performance in this regard include the use of so-called inverted configurations. By exciting surface plasmons from the back side of the grating, this design overcomes limitations of traditional grating-based SPR sensors caused by direct contact with the analyte medium and offers new design possibilities for implementing microfluidic analytical systems. Here a simulation for optimizing design parameters and a corresponding microfabrication technology to create the inverted grating are presented. An experimental evaluation of surface plasmon excitation and sensitivity enhancement demonstrates the feasibility of the approach. While the observed coupling effect of inverted grating structures is not as strong as the conventional configuration, it offers benefits like preventing surface oxidation, enabling repeated usage and new concepts for biosample processing.

Integration Of Acoustophoresis and Dielectrophoresis on A Single Chip Platform: A Numerical Study

Passive or active manipulation forces would be needed for the microfluidic system for chemical and biological analysis in order to regulate, trap, separate, sort, and discriminate between particles and cells. The primary goals of passive manipulation are consistency and repeatability to attain high levels of control with exact trajectories. In the meantime, by introducing external forces like hydrodynamic, dielectrophoretic, magnetophoretic, acoustophoretic, and optical tweezing, the active manipulations allow to control particle displacement in a highly predictable and consistent fashion. These methods are much more promising for the development of a small and compact biomedical diagnostic rapid test. Since most biological particles are suspended in different biological fluids like blood and urine, dielectrophoresis (DEP) and acoustophoresis (ACP) have been demonstrated to be promising among these external forces because of their ability to apply forces on the particles in a liquid environment. Additionally, both techniques are fast, inexpensive to fabricate, label-free, and incredibly selective. In this study, we introduce a novel method that combines these two forces into a single chip, improving the separation process for ACP and DEP based on the intrinsic dielectric and acoustic properties of the particles, respectively. It is anticipated that this research would shed light on why particular manipulative factors predominate more or less in particular situations.

A self-driven multifunctional microfluidic sweat analysis system for efficient sweat collection and real-time monitoring

Wearable sweat sensors offer the opportunity for non-invasive and real-time monitoring of metabolite information in sweat, enabling individuals to dynamically monitor their physiology states at the molecular level. However, sweat possesses characteristics such as small volume, volatility, susceptibility to epidermal contamination, and low analyte concentrations, making its reliable collection and detection challenge for wearable sweat sensors. In this work, we develop a self-driven multifunctional microfluidic sweat analysis system (SMMSAS) for efficient sweat collection and detection. The SMMSAS features with a wetting contrast for the sweat, which comprises superhydrophobic inlet, superhydrophilic microchannel enriched with nanofiber, and an absorbent test paper. Such a feature enables the spontaneous sweat transports in the form of dropwise due to the exertion of “Laplace force-capillary force-capillary force” on sweat, thereby facilitating the real-time sweat detection. SMMSAS integrated with the functions of impedance and colorimetry analysis is able to simultaneously record the multiple information in sweat, including sweat volume, sweating rate, total electrolyte concentration, glucose concentration, and pH value, all of which are important to reflect the physiological state of the human body. We also demonstrate the capability of SMMSAS in real-time monitoring the sweating conditions of the human body, which is attributed to its fast responsiveness (∼ 100 ms) for the wide range sweating rate (0.5−15 μL/min) and various total electrolyte concentration (0−80 mM), paving the way for next-generation personalized healthcare.

Establishment of a novel microfluidic co-culture system for simultaneous analysis of multiple indicators of gefitinib sensitivity in colorectal cancer cells.

The therapeutic effect of gefitinib on colorectal cancer (CRC) is unclear, but it has been reported that stromal cells in the tumor microenvironment may have an impact on drug sensitivity. Herein, we established a microfluidic co-culture system and explored the sensitivity of CRC cells co-cultured with cancer-associated fibroblasts (CAFs) to gefitinib. The system consisted of a multichannel chip and a Petri dish. The chambers in the chip and dish were designed to continuously supply nutrients for long-term cell survival and create chemokine gradients for driving cell invasion without any external equipment. Using this system, the proliferation and invasiveness of cells were simultaneously evaluated by quantifying the area of cells and the migration distance of cells. In addition, the system combined with live cell workstation could evaluate the dynamic drug response of co-cultured cells and track individual cell trajectories in real-time. When CRC cells were co-cultured with CAFs, CAFs promoted CRC cell proliferation and invasion and reduced the sensitivity of cells to gefitinib through the exosomes secreted by CAFs. Furthermore, the cells that migrated out of the chip were collected, and EMT-related markers were determined by immunofluorescent and western blot assays. The results demonstrated that CAFs affected the response of CRC cells to gefitinib by inducing EMT, providing new ideas for further research on the resistance mechanism of gefitinib. This suggests that targeting CAFs or exosomes might be a new approach to enhance CRC sensitivity to gefitinib, and our system could be a novel platform for investigating the crosstalk between tumor cells and CAFs and understanding multiple biological changes of the tumor cells in the tumor microenvironment.

Miniaturized Biosensors Based on Lanthanide-Doped Upconversion Polymeric Nanofibers.

Electrospun nanofibers possess a large surface area and a three-dimensional porous network that makes them a perfect material for embedding functional nanoparticles for diverse applications. Herein, we report the trends in embedding upconversion nanoparticles (UCNPs) in polymeric nanofibers for making an advanced miniaturized (bio)analytical device. UCNPs have the benefits of several optical properties, like near-infrared excitation, anti-Stokes emission over a wide range from UV to NIR, narrow emission bands, an extended lifespan, and photostability. The luminescence of UCNPs can be regulated using different lanthanide elements and can be used for sensing and tracking physical processes in biological systems. We foresee that a UCNP-based nanofiber sensing platform will open opportunities in developing cost-effective, miniaturized, portable and user-friendly point-of-care sensing device for monitoring (bio)analytical processes. Major challenges in developing microfluidic (bio)analytical systems based on UCNPs@nanofibers have been reviewed and presented.

Programmable droplet microfluidics for complex multistep bioassays

AbstractProgrammable droplet microfluidics (PDM) refers to a microfluidic device that integrates the functionality of a series of droplets with distinct spatial locations in a designated temporal order. PDM streamlines the intricate workflow of complex bioassays by enabling programmable and macroscopic droplet displacements, in which the droplets serve as reservoirs for reagents, microvalves for liquid insulation, and in some cases micropumps for mass transportation. As these droplets are intangible structures, the need for expensive microfabrication procedures is eliminated. Furthermore, the parallelization of the droplet series provides flexibility in controlling the throughput of the microfluidic analysis system. This paper provides a comprehensive review of PDMs enabled by various microfluidic mechanisms, including magnetism actuation, relative liquid displacement, capillary suction and sequential microdisplacement. Additionally, important applications of PDM systems for nucleic acid detection, immunoassay, drug testing, and sample recovery are also introduced. In conclusion, PDM demonstrates its potential as a highly advantageous tool for executing intricate multistep bioassays on a microfluidic platform. These technologies have exhibited superiority over their traditional counterparts in terms of size reduction, automation, and low sample/reagent consumption.

A Novel Micromixer That Exploits Electrokinetic Vortices Generated on a Janus Droplet Surface.

Micromixers play a crucial role as essential components in microfluidic analysis systems. This paper introduces a novel micromixer designed by harnessing electrokinetic vortices arising on the surface of a Janus droplet within a microchannel. The Janus droplet is characterized by different polarities of charges on its two sides (upstream part and downstream part). In the presence of a direct current electric field, the droplet's surface generates electroosmotic flows in opposite directions, resulting in the formation of vortices and facilitating solution mixing. Results from numerical simulations suggest that a better mixing performance of the micromixer is associated with both a higher absolute value of the zeta potential ratio between the downstream and upstream surfaces of the Janus droplet and a larger downstream surface area. Additionally, this study reveals that microchannel dimensions significantly influence the performance of the micromixer. Smaller microchannel widths and heights correspond to a larger mixing index for the micromixer. The micromixer presented in this study features a simple structure, easy fabrication, and holds promising application potential.

Establishment of bladder cancer spheroids and cultured in microfluidic platform for predicting drug response.

Cisplatin-containing combination chemotherapy has been used as the standard treatment for bladder cancer patients at advanced stage. However, nearly 50% of patients are nonresponders. To guide the selection of more effective chemotherapeutic agents, a bladder cancer spheroids microfluidic drug sensitivity analysis system was established in this study. Bladder cancer spheroids were established and successfully cultured in a customized microfluidic device to assess their response to different chemotherapeutic agents. The in vitro drug sensitivity results were also compared to patient-derived xenograft (PDX) models and clinical responses of patients. As a result, bladder cancer spheroids faithfully recapitulate the histopathological and genetic features of their corresponding parental tumors. Furthermore, the in vitro drug sensitivity outcomes of spheroids (n = 8) demonstrated a high level of correlation with the PDX (n = 2) and clinical response in patients (n = 2). Our study highlights the potential of combining bladder cancer spheroids and microfluidic devices as an efficient and accurate platform for personalized selection of chemotherapeutic agents.

Microfluidic characterization of single-cell biophysical properties and the applications in cancer diagnosis.

Single-cell biophysical properties play a crucial role in regulating cellular physiological states and functions, demonstrating significant potential in the fields of life sciences and clinical diagnostics. Therefore, over the last few decades, researchers have developed various detection tools to explore the relationship between the biophysical changes of biological cells and human diseases. With the rapid advancement of modern microfabrication technology, microfluidic devices have quickly emerged as a promising platform for single-cell analysis offering advantages including high-throughput, exceptional precision, and ease of manipulation. Consequently, this paper provides an overview of the recent advances in microfluidic analysis and detection systems for single-cell biophysical properties and their applications in the field of cancer. The working principles and latest research progress of single-cell biophysical property detection are first analyzed, highlighting the significance of electrical and mechanical properties. The development of data acquisition and processing methods for real-time, high-throughput, and practical applications are then discussed. Furthermore, the differences in biophysical properties between tumor and normal cells are outlined, illustrating the potential for utilizing single-cell biophysical properties for tumor cell identification, classification, and drug response assessment. Lastly, we summarize the limitations of existing microfluidic analysis and detection systems in single-cell biophysical properties, while also pointing out the prospects and future directions of their applications in cancer diagnosis and treatment.

Droplet Transportation on Liquid-Infused Asymmetrically Structured Surfaces by Mechanical Oscillation and Viscosity Control.

The transportation of droplets on solid surfaces has received significant attention owing to its importance in biochemical analysis and microfluidics. In this study, we propose a novel strategy for controlling droplet motion by combining an asymmetric structure and infused lubricating oil on a vibrating substrate. The transportation of droplets with volumes ranging from 10 to 90 μL was realized, and the movement speed could be adjusted from 1.45 to 10.87 mm/s. Typical droplet manipulations, including droplet transportation along a long trajectory and selective movement of multiple droplets, were successfully demonstrated. Through experimental exploration and theoretical analysis, we showed that the adjustment of droplet transport velocity involves an intricate interaction among the Ohnesorge number, droplet volume, and input amplitude. It can potentially be used for the more complex manipulation of liquid droplets in microfluidic and biochemical analysis systems.

Microfluidic flow injection analysis system for the electrochemical detection of dopamine using diazonium-grafted copper nanoparticles on multi-walled carbon nanotube-modified surfaces

In this proof-of-concept study, a microfluidic flow injection analysis (FIA) system was developed using multi-walled carbon nanotube-modified screen-printed carbon electrodes (CNTSPEs) that were modified with copper nanoparticles (CuNPs) following the electrodeposition of the diazonium salt of 4-aminothiophenol to form 4-thiophenol-conjugated CuNPs (CuNPs-CNTSPE). Transmission electron microscopy (TEM), atomic force microscopy (AFM), and scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS) were used to characterize the size of CuNPs, morphology and elemental analysis of CuNPs-CNTSPE, respectively. Using electrochemical impedance spectroscopy (EIS), the charge-transfer resistance (Rct) of CuNPs-CNTSPE was found to be 20-fold lower than that of CNTSPE. The CuNPs-CNTSPE displayed an oxidation peak for dopamine at −0.08 V which is ∼80 mV lower than the one detected using CNTSPE. The modified electrode was used in microfluidic flow injection analysis and offline systems for sensitive detection of dopamine (DA). The pH, flow rate, loop volume, concentration, and type of surfactant were all optimized for on-chip detection. Under the optimal conditions, using phosphate electrolyte solution (pH 6) containing 0.05% (w/v) Tween 20® as the carrier at a flow rate of 0.6 mL min−1 and a loop volume of 50 μL, the calibration curve was linear from 1.5 to 500 nM with a limit of detection of 0.33 nM. This technique was used for the successful detection of DA in real samples with recovery ranging from 96.5% to 103.8%. The microfluidic FIA system described here has the potential to be used as an electrochemical point-of-care device for rapid DA detection with high sensitivity and reproducibility.

Establishment of a cascaded microfluidic single cell analysis system for molecular and functional heterogeneity analysis of circulating tumor cells

Single cell heterogeneity analysis of circulating tumor cells (CTCs) is expected to reveal important tumor biological information for the evaluation and prediction of tumor stage and metastatic progression. Although various methods have been proposed to isolate and detect CTCs, consecutive CTCs sorting and in-situ multi-dimensional heterogeneity analysis in a cascaded system remain challenging. Here, a cascaded microfluidic chip was constructed, successively integrating a spiral structure for size-based removal of blood cells and a single cell array structure for the subsequent capture of CTCs, complementing the advantages of high throughput and high separation purity for both structures. Importantly, the molecular and functional heterogeneity analysis of CTCs can be in-situ realized and real-time observed at the single-cell level. Using this system, we captured CTCs and situ-analyzed the heterogeneity of single CTCs from different types of cancer patients, finding that mesenchymal phenotype is present in more CTCs at the advanced stage, and CTCs show active motility and division ability in metastatic patients. Therefore, the cascaded microfluidic chip offers an effective strategy for studying the molecular and functional heterogeneity of single CTCs, providing critical information for the evaluation of tumor progression and metastasis.

Articles of Significant Interest in This Issue.

Sidi Mabrouk et al. (e01596-22) adapted a high-throughput microfluidic system for long-term analysis of bacteriophage-bacterium interactions. The team applied the method to isolate phages from environmental samples. The use of an off-the-shelf product for phage studies is important to advance fundamental science in the field and facilitate environmental surveillance work.

Regulating the Gas–Liquid Slug Flow in Microchannels through High-Frequency Pulsatile Perturbations

The gas–liquid slug flow that is characterized with alternate formation of gas and liquid segments has been widely applied for chemical synthesis and analysis. For its applications, it is essential to precisely control the gas and liquid slug lengths. However, using the passive method, the bubble/droplet generation frequency and their sizes are influenced by multiple parameters including the channel dimension, fluid physical properties, the gas-to-liquid flow rate ratio, and so forth and hence lack control flexibility. In this work, an active approach using pulsatile perturbations to control the gas–liquid slug flow is demonstrated. A rotating valve is applied to produce a high-frequency pulsating air flow. Under its influence, either Taylor bubbles in a hydrophilic channel or liquid-in-gas droplets in a hydrophobic channel can be generated synchronously with the flow pulsation. Therefore, the gas and liquid slug lengths can be independently tuned via the air inlet pressure and the liquid flow rate, respectively. The maximum generation frequency is up to ∼1300 Hz. In addition, the effects of the pulsation frequency, the pulse duty ratio, and the liquid viscosity are also investigated. This technique provides an efficient method to regulate the gas–liquid slug flow in multiphase microreactors or microfluidic analytical systems.

Optical Technologies for Single-Cell Analysis on Microchips

Cell analysis at the single-cell level is of great importance to investigate the inherent heterogeneity of cell populations and to understand the morphology, composition, and function of individual cells. With the continuous innovation of analytical techniques and methods, single-cell analysis on microfluidic chip systems has been extensively applied for its precise single-cell manipulation and sensitive signal response integrated with various detection techniques, such as optical, electrical, and mass spectrometric analyses. In this review, we focus on the specific optical events in single-cell analysis on a microfluidic chip system. First, the four most commonly applied optical technologies, i.e., fluorescence, surface-enhanced Raman spectroscopy, surface plasmon resonance, and interferometry, are briefly introduced. Then, we focus on the recent applications of the abovementioned optical technologies integrated with a microfluidic chip system for single-cell analysis. Finally, future directions of optical technologies for single-cell analysis on microfluidic chip systems are predicted.

Recent Progress and Applications of Thermal Lens Spectrometry and Photothermal Beam Deflection Techniques in Environmental Sensing.

This paper presents recent development and applications of thermal lens microscopy (TLM) and beam deflection spectrometry (BDS) for the analysis of water samples and sea ice. Coupling of TLM detection to a microfluidic system for flow injection analysis (μFIA) enables the detection of microcystin-LR in waters with a four samples/min throughput (in triplicate injections) and provides an LOD of 0.08 µg/L which is 12-times lower than the MCL for microcystin-LR in water. μFIA-TLM was also applied for the determination of total Fe and Fe(II) in 3 µL samples of synthetic cloudwater. The LODs were found to be 100 nM for Fe(II) and 70 nM for total Fe. The application of µFIA-TLM for the determination of ammonium in water resulted in an LOD of 2.3 µM for injection of a 5 µL sample and TLM detection in a 100 µm deep microfluidic channel. For the determination of iron species in sea ice, the BDS was coupled to a diffusive gradient in the thin film technique (DGT). The 2D distribution of Fe(II) and total Fe on DGT gels provided by the BDS (LOD of 50 nM) reflected the distribution of Fe species in sea ice put in contact with DGT gels.

Integrated "lab-on-a-chip" microfluidic systems for isolation, enrichment, and analysis of cancer biomarkers.

The liquid biopsy has garnered considerable attention as a complementary clinical tool for the early detection, molecular characterization and monitoring of cancer over the past decade. In contrast to traditional solid biopsy techniques, liquid biopsy offers a less invasive and safer alternative for routine cancer screening. Recent advances in microfluidic technologies have enabled handling of liquid biopsy-derived biomarkers with high sensitivity, throughput, and convenience. The integration of these multi-functional microfluidic technologies into a 'lab-on-a-chip' offers a powerful solution for processing and analyzing samples on a single platform, thereby reducing the complexity, bio-analyte loss and cross-contamination associated with multiple handling and transfer steps in more conventional benchtop workflows. This review critically addresses recent developments in integrated microfluidic technologies for cancer detection, highlighting isolation, enrichment, and analysis strategies for three important sub-types of cancer biomarkers: circulating tumor cells, circulating tumor DNA and exosomes. We first discuss the unique characteristics and advantages of the various lab-on-a-chip technologies developed to operate on each biomarker subtype. This is then followed by a discussion on the challenges and opportunities in the field of integrated systems for cancer detection. Ultimately, integrated microfluidic platforms form the core of a new class of point-of-care diagnostic tools by virtue of their ease-of-operation, portability and high sensitivity. Widespread availability of such tools could potentially result in more frequent and convenient screening for early signs of cancer at clinical labs or primary care offices.