Hand-Powered Point-of-Care: Centrifugal Microfluidic Platform for Urine Routine Examination (µCUREX).

Urinalysis is one of the simplest and most common medical tests in modern cities. With the assistance of professional technicians and equipment, people in metropolitan areas can effortlessly acquire information about their physiological conditions from traditional clinical laboratories. However, the threshold, including precise benchtop equipment and well-trained personnel, still remains a considerable dilemma for residents in healthcare-poor areas. Hence, it is a crucial and urgent topic to develop a smart and affordable widget to address this challenge. To improve the healthcare rights of residents, we proposed a disposable centrifugal microfluidic urine routine examination platform (named µCUREX) actuated with a modified hand-powered fan. Two parts of urinalysis (sediment test and chemical strip test) were integrated into the µCUREX disc. The influence on sedimentation by variant hand-powered manipulation was simulated using COMSOL. As a result, more than 70% of the sediment can be collected. Moreover, the color change of chemical strip papers (indicators for glucose, pH, protein, and occult blood) was recorded with a 3D-printed studio and analyzed after reaction with chemical-spiked and pH-adjusted artificial and human urine specimens. The whole process can be completed within 10 min, with only 200 µL of urine needed. In conclusion, we successfully constructed an ultra-low-cost point-of-care platform for urinalysis in extremely resource-poor settings. The handy size, high affordability, and user-friendliness of the µCUREX disc provide strong potential and feasibility in solving problems in resource-poor settings. Furthermore, we highly expect the µCUREX platform to improve the level of healthcare in resource-limited areas.

Antifouling Properties of Amine-Oxide-Containing Zwitterionic Polymers.

Biofouling due to nonspecific proteins or cells on the material surfaces is a major challenge in a range of applications such as biosensors, medical devices, and implants. Even though poly(ethylene glycol) (PEG) has become the most widely used stealth material in medical and pharmaceutical products, the number of reported cases of PEG-triggered rare allergic responses continues to increase in the past decades. Herein, a new type of antifouling material poly(amine oxide) (PAO) has been evaluated as an alternative to overcome nonspecific foulant adsorption and impart comparable biocompatibility. Alkyl-substituted PAO containing diethyl, dibutyl, and dihexyl substituents are prepared, and their solution properties are studied. Photoreactive copolymers containing benzophenone as the photo-cross-linker are prepared by reversible addition-fragmentation chain-transfer polymerization and fully characterized by gel permeation chromatography and dynamic light scattering. Then, these water-soluble polymers are anchored onto a silicon wafer with the aid of UV irradiation. By evaluating the fouling resistance properties of these modified surfaces against various types of foulants, protein adsorption and bacterial attachment assays show that the cross-linked PAO-modified surface can efficiently inhibit biofouling. Furthermore, human blood cell adhesion experiments demonstrate that our PAO polymer could be used as a novel surface modifier for biomedical devices.

Antifouling strategies in advanced electrochemical sensors and biosensors.

Electrochemical biosensors have been applied in a broad range of clinical applications for pathogen biomarker detection and medical applications and diagnosis due to the sensitivity of electrochemical methods and the bioselectivity of the components. The complexity of clinical conditions with various biofoulants (proteins, cells, polysaccharides and lipids) severely influences the reliability and stability of sensors for direct detection or immersion under changing conditions. Therefore, designing an antifouling sensing platform that can effectively reduce undesired binding to maintain biosensor performance in optimized analysis is necessary. For this purpose, the fundamental mechanisms of fouling materials and commonly used biocompatible antifouling components have been discussed, and the relevant effective modification strategies are introduced in this review. Recent advances in these strategies are demonstrated in examples with analysis of essential modification methods for reliable sensing in non-specific binding solutions or complex biofluids. The challenges and future perspectives of modification strategies for current clinical application are also discussed in this review.

Effective Construction of a High-Capacity Boronic Acid Layer on a Quartz Crystal Microbalance Chip for High-Density Antibody Immobilization.

Boronic acids (BAs) provide strong potential in orientation immobilization of antibody and the modification method is crucial for efficiency optimization. A highly effective method has been developed for rapid antibody immobilization on gold electrodes through the electrodeposition of a BA⁻containing linker in this study. Aniline-based BA forms a condense layer while antibody could automatically immobilize on the surface of the electrode. Compare to traditional self-assembled monolayer method, the electrodeposition process dramatically reduces the modification time from days to seconds. It also enhances the immobilized efficiency from 95 to 408 (ng/cm²) with a strong preference being exhibited for shorter aniline-based linkers.

Noninvasive Glucose Monitoring with a Contact Lens and Smartphone.

Diabetes has become a chronic metabolic disorder, and the growing diabetes population makes medical care more important. We investigated using a portable and noninvasive contact lens as an ideal sensor for diabetes patients whose tear fluid contains glucose. The key feature is the reversible covalent interaction between boronic acid and glucose, which can provide a noninvasive glucose sensor for diabetes patients. We present a phenylboronic acid (PBA)-based HEMA contact lens that exhibits a reversible swelling/shrinking effect to change its thickness. The difference in thickness can be detected in a picture taken with a smartphone and analyzed using software. Our novel technique offers the following capabilities: (i) non-enzymatic and continuous glucose detection with the contact lens; (ii) no need for an embedded circuit and power source for the glucose sensor; and (iii) the use of a smartphone to detect the change in thickness of the contact lens with no need for additional photo-sensors. This technique is promising for a noninvasive measurement of the glucose level and simple implementation of glucose sensing with a smartphone.

Advances in nanowire transistors for biological analysis and cellular investigation.

Electrical biosensors based on silicon nanowire field-effect transistors (SiNW-FETs) have attracted enormous interest in the biosensing field. SiNW-FETs have proven to be significant and efficient in detecting diverse biomolecular species with the advantages of high probing sensitivity, target selectivity, real-time recording and label-free detection. In recent years, significant advances in biosensors have been achieved, particularly for cellular investigation and biomedical diagnosis. In this critical review, we will report on the latest developments in biosensing with SiNW-FETs and discuss recent advancements in the innovative designs of SiNW-FET devices. This critical review introduces the basic instrumental setup and working principle of SiNW-FETs. Technical approaches that attempted to enhance the detection sensitivity and target selectivity of SiNW-FET sensors are discussed. In terms of applications, we review the recent achievements with SiNW-FET biosensors for the investigations of protein-protein interaction, DNA/RNA/PNA hybridization, virus detection, cellular recording, biological kinetics, and clinical diagnosis. In addition, the novel architecture designs of the SiNW-FET devices are highlighted in studies of live neuron cells, electrophysiological measurements and other signal transduction pathways. Despite these remarkable achievements, certain improvements remain necessary in the device performance and clinical applications of FET-based biosensors; thus, several prospects about the future development of nanowire transistor-based instruments for biosensing employments are discussed at the end of this review.

Development of a thermotaxis and rheotaxis microfluidic device for motile spermatozoa sorting.

Male infertility is a pervasive global reproductive challenge, primarily attributed to a decline in semen quality. Addressing this concern, there has been a growing focus on spermatozoa sorting in assisted reproductive technology. This study introduces a groundbreaking development in the form of a thermotaxis and rheotaxis microfluidic (TRMC) device designed for efficient motile spermatozoa sorting within a short 15-min timeframe. The TRMC device mimics the natural sperm sorting mechanism of the oviduct, selecting spermatozoa with superior motility and DNA integrity. The experimental outcomes demonstrate a remarkable enhancement in the percentage of progressive spermatozoa following sorting, soaring from 3.90% to an impressive 96.11% when subjected to a temperature decrease from 38 °C to 35 °C. Notably, sperm motility exhibited a substantial 69% improvement. The TRMC device exhibited a commendable recovery rate of 60.93%, surpassing current clinical requirements. Furthermore, the sorted spermatozoa displayed a notable reduction in the DNA fragmentation index to 6.94%, signifying a substantial 90% enhancement in DNA integrity. This remarkable advancement positions the TRMC device as highly suitable for applications in in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI), offering a promising solution to male infertility challenges.

An ultrasensitive nanowire-transistor biosensor for detecting dopamine release from living PC12 cells under hypoxic stimulation.

Dopamine (DA) is an important neurotransmitter that is involved in neuronal signal transduction and several critical illnesses. However, the concentration of DA is extremely low in patients and is difficult to detect using existing electrochemical biosensors with detection limits typically around nanomolar levels (∼10(-9) M). Here, we developed a nanoelectronic device as a biosensor for ultrasensitive and selective DA detection by modifying DNA-aptamers on a multiple-parallel-connected (MPC) silicon nanowire field-effect transistor (referred to as MPC aptamer/SiNW-FET). Compared with conventional electrochemical methods, the MPC aptamer/SiNW-FET has been demonstrated to improve the limit of DA detection to <10(-11) M and to possess a detection specificity that is able to distinguish DA from other chemical analogues, such as ascorbic acid, catechol, phenethylamine, tyrosine, epinephrine, and norepinephrine. This MPC aptamer/SiNW-FET was also applied to monitor DA release under hypoxic stimulation from living PC12 cells. The real-time recording of the exocytotic DA induced by hypoxia reveals that the increase in intracellular Ca(2+) that is required to trigger DA secretion is dominated by an extracellular Ca(2+) influx, rather than the release of intracellular Ca(2+) stores.

Wearable Microfluidics for Continuous Assay.

The development of wearable devices provides approaches for the realization of self-health care. Easily carried wearable devices allow individual health monitoring at any place whenever necessary. There are various interesting monitoring targets, including body motion, organ pressure, and biomarkers. An efficient use of space in one small device is a promising resolution to increase the functions of wearable devices. Through integration of a microfluidic system into wearable devices, embedding complicated structures in one design becomes possible and can enable multifunction analyses within a limited device volume. This article reviews the reported microfluidic wearable devices, introduces applications to different biofluids, discusses characteristics of the design strategies and sensing principles, and highlights the attractive configurations of each device. This review seeks to provide a detailed summary of recent advanced microfluidic wearable devices. The overview of advanced key components is the basis for the development of future microfluidic wearable devices.

Rapid detection of live bacteria in water using nylon filter membrane-integrated centrifugal microfluidics.

Water is one of the most indispensable elements for human beings. People can live without food for a couple of weeks but cannot live without water for a couple of days. Unfortunately, drinking water is not always safe around the world; in many areas, the water for drinking could be contaminated with various microbes. However, the total viable microbe count in water still relies on culture-based methods in laboratories. Therefore, in this work, we report a novel, simple, and highly efficient strategy to detect live bacteria in water via a nylon membrane-integrated centrifugal microfluidic device. A handheld fan and a rechargeable hand warmer were utilized as the centrifugal rotor and the heat resource for reactions, respectively. The bacteria in water can be rapidly concentrated >500-fold by our centrifugation system. After incubation with water-soluble tetrazolium-8 (WST-8), the color change of the nylon membranes can be visually interpreted directly by the naked eye or recorded with a smartphone camera. The whole process can be finished in 3 h, and the detection limit can reach 102 CFU/mL. The detection range ranges from 102 CFU/mL to 105 CFU/mL. The cell counting results of our platform are highly positively correlated with the results of cell counting by the conventional lysogeny broth (LB) agar plate approach or the commercial 3 M Petrifilm™ cell counting plate. Our platform provides a convenient and sensitive strategy for rapid monitoring. We highly anticipate that this platform can improve water quality monitoring in resource-poor countries in the near future.

Application of aminobenzoic acid electrodeposited screen-printed carbon electrode in the beta-amyloid electrochemical impedance spectroscopy immunoassay.

Alzheimer's disease (AD) is one of the important neurodegenerative diseases, in the modern aging society, it has become an issue people need to work on. Of the pathogenic factor which leads to AD, beta-amyloid (Aß) is the most important one. It can form the senile plaque which aggregates in the neuron and interrupts the signal transmission. This research is based on the electrochemical system and screen-printed carbon electrode (SPCE) incorporated with pretreatment, electrodeposition, electrochemical impedance spectroscopy (EIS), antibody, and blocking agent. This immunosensor is applied to detect the different concentrations of Aß. The standard curve between electrical impedance and concentration of Aß is calculated. The specificity of the immunosensor is tested. This survey optimizes the electrodeposition condition for 4-aminobenzoic acid (4-ABA) and the parameter for antibody and blocking agents. This study fabricates a more dense, uniform, and stable film of 4-ABA. This sensor presents a range of detection from 1 fg/ml to 100 pg/ml and a limit of detection to 3.84 fg/ml. This sensor can identify the isoform of Aß. This research shortens the fabricating time to 3.5 h. This study fabricates a label-free and low-cost immunosensor for Aß with a short fabricating time, high stability, wide range of detection, low limit of detection, and good specificity. The impedance of the carbon printed electrodes is very high and is always measured by its current but this study provides a fabrication technique for high-efficiency carbon printed electrodes for electrochemical impedance spectroscopy sensing.

Design of a gradient-rheotaxis microfluidic chip for sorting of high-quality Sperm with progressive motility.

Assisted reproductive technology (ART) is an important invention for the treatment of human infertility, and the isolation of high-quality sperm with progressive motility is one of the most critical steps that eventually affect the fertilization rate. Conventional sperm separation approaches include the swim-up method and density gradient centrifugation. However, the quality of isolated sperm obtained from both approaches can still be improved by improving sorted sperm motility, minimizing the DNA fragmentation rate, and removing abnormal phenotypes. Here, we report a Progressive Sperm Sorting Chip (PSSC) for high-quality sperm isolation. Based on the rheotaxis behavior of sperm, a gradient flow field is created in the chip for progressive sperm sorting. Clinical experiment results for 10 volunteers showed that greater than 90% of isolated sperm exhibit high motility (> 25 µm/s), high linearity (0.8), and a very low DNA fragmentation rate (< 5%). In addition, the whole process is label and chemical free. These features aid in gentle sperm sorting to obtain healthy sperm. This device uniquely enables the selection of high-quality sperm with progressive motility and might be clinically applied for infertility treatment in the near future.

Wearable hydrogel patch with noninvasive, electrochemical glucose sensor for natural sweat detection.

Recent advances in microelectronics and electrochemical sensing platforms have preceded the development of devices for personal monitoring and managing physiological and metabolic information that exploit sweat as a noninvasive, convenient approach for providing information about underlying health conditions, such as glucose level monitoring. Although most sweat glucose sensors have targeted applications during exercise and other active stimulation induced-sweat, natural sweating offers an attractive alternative with minimal effect on users that can be accessed during routine and sedentary activities without impeding personal lifestyle and preserves the correlation between blood and sweat glucose. Here, we present a noninvasive sweat glucose sensor with convenient hydrogel patches for rapid sampling of natural perspiration without external activities that stimulate sweating. The wearable hydrogel patch rapidly takes up natural sweat from the hand and serves as a medium for electrochemical sensing. A prussian blue-doped poly(3,4-ethylenedioxythiophene nanocomposite (PB-PEDOT NC) electrode provides cost-effective, stable and excellent electrocatalytic activity in sweat glucose measurements. We demonstrated sweat glucose sensor functionality by long-term measurements of glucose in sweat from human subjects consuming food and drinks. By enabling the analysis of sweat glucose during routine and sedentary activities, the sweat glucose sensor shows great promise for clinical-grade glucose management and enlarges the scope of next-generation noninvasive sensing systems.

Passively driven microfluidic device with simple operation in the development of nanolitre droplet assay in nucleic acid detection.

Since nucleic acid amplification technology has become a vital tool for disease diagnosis, the development of precise applied nucleic acid detection technologies in point-of care testing (POCT) has become more significant. The microfluidic-based nucleic acid detection platform offers a great opportunity for on-site diagnosis efficiency, and the system is aimed at user-friendly access. Herein, we demonstrate a microfluidic system with simple operation that provides reliable nucleic acid results from 18 uniform droplets via LAMP detection. By using only micropipette regulation, users are able to control the nanoliter scale of the droplets in this valve-free and pump-free microfluidic (MF) chip. Based on the oil enclosure method and impermeable fabrication, we successfully preserved the reagent inside the microfluidic system, which significantly reduced the fluid loss and condensation. The relative standard deviation (RSD) of the fluorescence intensity between the droplets and during the heating process was < 5% and 2.0%, respectively. Additionally, for different nucleic acid detection methods, the MF-LAMP chip in this study showed good applicability to both genome detection and gene expression analysis.

Rapid construct superhydrophobic microcracks on the open-surface platform for droplet manipulations.

Droplet-based transport driven by surface tension has been explored as an automated pumping source for several biomedical applications. This paper presented a simple and fast superhydrophobic modify and patterning approach to fabricate various open-surface platforms to manipulate droplets to achieve transport, mixing, concentration, and rebounding control. Several commercial reagents were tested in our approach, and the Glaco reagent was selected to create a superhydrophobic layer; laser cutters are utilized to scan on these superhydrophobic surface to create gradient hydrophilic micro-patterns. Implementing back-and-forth vibrations on the predetermined parallel patterns, droplets can be transported and mixed successfully. Colorimetry of horseradish peroxidase (HRP) mixing with substrates also reduced the reaction time by more than 5-times with the help of superhydrophobic patterned chips. Besides, patterned superhydrophobic chips can significantly improve the sensitivity of colorimetric glucose-sensing by more than 10 times. Moreover, all bioassays were distributed homogeneously within the region of hydrophilic micropatterns without the coffee-ring effect. In addition, to discuss further applications of the surface wettability, the way of controlling the droplet impacting and rebounding phenomenon was also demonstrated. This work reports a rapid approach to modify and patterning superhydrophobic films to perform droplet-based manipulations with a lower technical barrier, higher efficiency, and easier operation. It holds the potential to broaden the applications of open microfluidics in the future.

Integration of Ni/NiO nanoparticles and a microfluidic ELISA chip to generate a sensing platform for Streptococcus pneumoniae detection.

Enzyme-linked immunosorbent assays (ELISAs) are tests that uses antibody recognition and enzyme catalytic activity to identify a substance, and they have been widely used as a diagnostic tool in the clinic. However, performing an ELISA requires various liquid handling steps and long binding times. To solve this problem, we developed a magnetic microfluidic ELISA system (MMF-ELISA). Integration with nickel magnetic nanoparticles can streamline the ELISA process in a fully automated manner for Streptococcus pneumoniae detection. First, we synthesized paramagnetic surface-oxidized nickel nanoparticles (Ni/NiO NPs) to carry protein G. Then, we assembled a SUM290 (UlaG)-specific antibody on protein G. Finally, we integrated the NPs on a microfluidics chip for S. pneumoniae detection. The chip contains three different layers to trap the solutions; the bottom layer SiO2 is patterned on hydrophobic polymers and integrated with the middle layer PDMS and the top layer PMMA. With Arduino and motor IC, we developed an automated platform for S. pneumoniae detection. Microfluidic ELISAs can reduce the manual handling and operation time. Furthermore, the developed system can be extended to multiple areas for ELISA-related assays. This economical, rapid and portable system may become a promising platform for sensing S. pneumoniae in clinical applications.

Construction of the Nickel Oxide Nanocoral Structure on Microscope Slides for Total Self-Assembly-Oriented Probe Immobilization and Signal Enhancement.

Proper orientation of probes and the binding capacity of surfaces will determine the performance of bio-applications. It has been reported that immobilizing through bio-/chemical affinity is an efficient but gentle strategy to solve the above-mentioned issue. Herein, we introduce a total self-assembly approach via the strong affinity of nickel oxide (NiO) to the polyhistidine-tag (His-tag). It allows the efficient immobilizing His-tagged proteins with orientation. Furthermore, we find that the nanocoral structure can be formed after applying rapid thermal annealing at 1100 °C, which could increase the His-tagged protein binding capacity efficiently by the enhanced surface-to-volume ratio. Lastly, we demonstrate the NiO thin film with the nanocoral structure, which has great potential for universal biosensing with a wide range of biomolecules, including DNA, protein, and bacteria. Through His-tagged monomer streptavidin (His6-mSA) or His-tagged protein G (His6-protein G), the biotinylated DNA or antibody could be immobilized with proper orientation on the surface consequently to complete a sensitive biomolecule detection. Moreover, the NiO nanocoral structure has the advantages of high hydrophilicity, transmittance, and pH stability that are promising to develop into several kinds of bio-applications in the near future.

A Noninvasive Wearable Device for Real-Time Monitoring of Secretion Sweat Pressure by Digital Display.

Sweat-based wearable devices have attracted increasing attention by providing abundant physiological information and continuous measurement through noninvasive healthcare monitoring. Sweat pressure generated via sweat glands to the skin surface associated with osmotic effects may help to elucidate such parameters as physiological conditions and psychological factors. This study introduces a wearable device for measuring secretion sweat pressure through noninvasive, continuous monitoring. Secretion pressure is detected by a microfluidic chip that shows the resistance variance from a paired electrode pattern and transfers digital signals to a smartphone for real-time display. A human study demonstrates this measurement with different exercise activities, showing the pressure ranges from 1.3 to 2.5 kPa. This device is user-friendly and applicable to exercise training and personal health care. The convenience and easy-to-wear characteristics of this device may establish a foundation for future research investigating sweat physiology and personal health care.

An antifouling peptide-based biosensor for determination of Streptococcus pneumonia markers in human serum.

We report a peptide-based sensor that involves a multivalent interaction with L-ascorbate 6-phosphate lactonase (UlaG), a protein marker of Streptococcus pneumonia. By integrating the antifouling feature of the sensor, we significantly improved the signal-to-noise ratio of UlaG detection. The antifouling surface was fabricated via electrodeposition using an equivalent mixture of 4-amino-N,N,N-trimethylanilinium and 4-aminobenzenesulfonate. This antifouling layer not only effectively reduces the non-specific adsorption on the biosensor but also decreases the charge transfer resistance (Rct) of the screen-printed carbon electrode. The aniline-modified S7 peptide, an UlaG-binding peptide, was pre-synthesized and further electrochemically modified to bind onto the antifouling layer. Bio-electrochemical analysis confirms that the antifouling S7-peptide sensor binds strongly to the UlaG with a dissociation constant (Kd) = 0.5 nM. This strong interaction can be attributed to a multivalent interaction between the biosensor and the heximeric form of UlaG. To demonstrate the potential for clinical application, further detection of Streptococcus pneumonia from 50 to 5×104 CFU/mL were successfully performed in 25% human serum.

Rapid purification of lung cancer cells in pleural effusion through spiral microfluidic channels for diagnosis improvement.

Lung cancer is one of the leading causes of death worldwide. Fifteen percent of lung cancer patients will present with malignant pleural effusion initially, and up to 50% will have malignant pleural effusion throughout the course of the disease. In this study, we developed a spiral microfluidic device that can rapidly isolate cancer cells and improve their purity through fluid dynamics. This label-free, high-throughput device continuously isolates cancer cells and other unrelated molecules from pleural effusion. Most of the background cells that affect interpretation are flushed to outlets 1 to 3, and cancer cells are hydrodynamically concentrated to outlet 4, with 90% of lung cancer cells flowing to this outlet. After processing, the purity of cancer cells identified in pleural effusion by CD45 and epithelial cell adhesion molecule (EpCAM) antibodies in flow cytometry will be increased by 6 to 24 times. The microfluidic device presented here has the advantages of rapid processing and low cost, which are conducive to rapid and accurate clinical diagnosis.