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Covalent organic frameworks (COFs) are a novel family of porous crystalline materials utilized in various advanced applications. However, applying COFs as a hazardous organic acid gas sensor is substantial but still challenging. Herein, a phenylenediamine-based covalent organic framework (TPDA-TPB COF) featuring excellent crystallinity, ultrastable thermal stability, and high surface area was successfully constructed. Then, the TPDA-TPB COF-modified quartz crystal microbalance (QCM) sensor is fabricated by immobilizing the TPDA-TPB COF thin film on the gold-QCM chip. The fabricated TPDA-TPB COF-modified QCM sensor demonstrates a rapid response, excellent reproducibility, high selectivity, and sensitivity to formic gas, arising from hydrogen-bonding interactions between formic acid and the outermost layer of the TPDA-TPB COF, as determined by extensive analysis and density functional theory calculations. The basic sites of the TPDA-TPB COF, which are numerous due to its high nitrogen content, and the carboxylic acid groups present in formic acid exhibit efficient interactions. The sensitivity of the TPDA-TPB COF-modified QCM sensor was found to be 7.75 Hzâ¯ppm-1 at standard room temperature and pressure conditions, with a limit of detection (LOD) of formic acid down to 1.18 ppm, which is significantly below the workplace olfactory threshold limit of 5.0 ppm established by the Occupational Safety and Health Administration. The TPDA-TPB COF-modified QCM sensor exhibits remarkable detecting capabilities, making it highly attractive for detecting organic acid vapors in diverse applications that require superior performance.
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Electrochemical affinity biosensors have the potential to facilitate the development of multiplexed point-of-care diagnostics in complex biological fluids. However, their commercial viability has been hindered by challenges such as electrode biofouling and the lack of inherent redox properties. To address this unmet need, we have developed a universal nanocomposite coating which is unique in its ability to not only allow oriented conjugation of the biorecognition element but also specific detection directly in complex biological fluids like serum and urine owing to its built-in antifouling and redox capabilities, thus improving suitability for point of care testing. This multifunctional coating comprises a 3D porous crosslinked bovine serum albumin matrix for oriented conjugation and antifouling properties with embedded graphene nanosheets modified with amino-ferrocene for enhanced conductivity and mediator-free biosensing. The coating showed minimal signal degradation despite prolonged exposure to 1% bovine serum albumin, artificial urine and untreated human serum for up to 30 days. To demonstrate its utility, we fabricated and tested proof-of-concept electrochemical immunosensors for bladder cancer protein biomarkers, specifically interleukin-8 (IL-8) and vascular endothelial growth factor (VEGF). The practical feasibility was highlighted by the excellent sensitivity and specificity observed for IL-8 and VEGF with a limit of detection of 41 pg mL-1 and 67 pg mL-1, respectively. Consequently, this universal nanocomposite-based electrochemical biosensing platform can be extended to the point of care testing of a broad spectrum of biomarkers present in complex biological fluids, thus enabling reliable and early diagnostics.
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Técnicas Biossensoriais , Técnicas Eletroquímicas , Grafite , Metalocenos , Nanocompostos , Oxirredução , Soroalbumina Bovina , Técnicas Biossensoriais/métodos , Nanocompostos/química , Humanos , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/instrumentação , Grafite/química , Soroalbumina Bovina/química , Fator A de Crescimento do Endotélio Vascular/sangue , Fator A de Crescimento do Endotélio Vascular/urina , Interleucina-8/sangue , Interleucina-8/urina , Interleucina-8/análise , Incrustação Biológica/prevenção & controle , Animais , Neoplasias da Bexiga Urinária/urina , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/urina , Compostos Ferrosos/química , BovinosRESUMO
In this study, we investigated a novel approach to fabricate multifunctional ionic gel sensors by using deep eutectic solvents (DESs) as replacements for water. When two distinct DESs were combined, customizable mechanical and conductive properties were created, resulting in improved performance compared with traditional hydrogel-based strain sensors. DES ionic gels possess superior mechanical properties, transparency, biocompatibility, and antimicrobial properties, making them suitable for a wide range of applications such as flexible electronics, soft robotics, and healthcare. We conducted a comprehensive evaluation of the DES ionic gels, evaluating their performance under extreme temperature conditions (-70 to 80 °C), impressive optical transparency (94%), and biocompatibility. Furthermore, a series of tests were conducted to evaluate the antibacterial performance (Escherichia coli) of the DES ionic gels. Their wide strain (1-400%) and temperature (15-50 °C)-sensing ranges demonstrate the versatility and adaptability of DES ionic gels for diverse sensing requirements. The resulting DES ionic gels were successfully applied in human activity and vital sign monitoring, demonstrating their potential for biointegrated sensing devices and healthcare applications. This study offers valuable insights into the development and optimization of hydrogel sensors, particularly for applications that require environmental stability, biocompatibility, and antibacterial performance, thereby paving the way for future advancements in this field.
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Antibacterianos , Solventes Eutéticos Profundos , Humanos , Solventes , Antibacterianos/farmacologia , Hidrogéis/farmacologia , Água , Escherichia coli , ÍonsRESUMO
This work focuses on porous organic polymers (POPs), which have gained significant global attention for their potential in energy storage and carbon dioxide (CO2) capture. The study introduces the development of two novel porous organic polymers, namely FEC-Mel and FEC-PBDT POPs, constructed using a simple method based on the ferrocene unit (FEC) combined with melamine (Mel) and 6,6'-(1,4-phenylene)bis(1,3,5-triazine-2,4-diamine) (PBDT). The synthesis involved the condensation reaction between ferrocenecarboxaldehyde monomer (FEC-CHO) and the respective aryl amines. Several analytical methods were employed to investigate the physical characteristics, chemical structure, morphology, and potential applications of these porous materials. Through thermogravimetric analysis (TGA), it was observed that both FEC-Mel and FEC-PBDT POPs exhibited exceptional thermal stability. FEC-Mel POP displayed a higher surface area and porosity, measuring 556 m2 g-1 and 1.26 cm3 g-1, respectively. These FEC-POPs possess large surface areas, making them promising materials for applications such as supercapacitor (SC) electrodes and gas adsorption. With 82 F g-1 of specific capacitance at 0.5 A g-1, the FEC-PBDT POP electrode has exceptional electrochemical characteristics. In addition, the FEC-Mel POP showed remarkable CO2 absorption capabilities, with 1.34 and 1.75 mmol g-1 (determined at 298 and 273 K; respectively). The potential of the FEC-POPs created in this work for CO2 capacity and electrical testing are highlighted by these results.
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Dióxido de Carbono , Polímeros , Metalocenos , PorosidadeRESUMO
In this study, we synthesized two conjugated microporous polymers (CMPs), An-Ph-TPA and An-Ph-Py CMPs, using the Suzuki cross-coupling reaction. These CMPs are organic polymers with p-conjugated skeletons and persistent micro-porosity and contain anthracene (An) moieties linked to triphenylamine (TPA) and pyrene (Py) units. We characterized the chemical structures, porosities, thermal stabilities, and morphologies of the newly synthesized An-CMPs using spectroscopic, microscopic, and N2 adsorption/desorption isotherm techniques. Our results from thermogravimetric analysis (TGA) showed that the An-Ph-TPA CMP displayed better thermal stability with Td10 = 467 °C and char yield of 57 wt% compared to the An-Ph-Py CMP with Td10 = 355 °C and char yield of 54 wt%. Furthermore, we evaluated the electrochemical performance of the An-linked CMPs and found that the An-Ph-TPA CMP had a higher capacitance of 116 F g-1 and better capacitance stability of 97% over 5000 cycles at 10 A g-1. In addition, we assessed the biocompatibility and cytotoxicity of An-linked CMPs using the MTT assay and a live/dead cell viability assay and observed that they were non-toxic and biocompatible with high cell viability values after 24 or 48 h of incubation. These findings suggest that the An-based CMPs synthesized in this study have potential applications in electrochemical testing and the biological field.
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Aminas , Polímeros , Polímeros/química , Adsorção , AntracenosRESUMO
Porous organic polymers (POPs) have plenteous exciting features due to their attractive combination of microporosity with π-conjugation. Nevertheless, electrodes based on their pristine forms suffer from severe poverty of electrical conductivity, precluding their employment within electrochemical appliances. The electrical conductivity of POPs may be significantly improved and their porosity properties could be further customized by direct carbonization. In this study, we successfully prepared a microporous carbon material (Py-PDT POP-600) by the carbonization of Py-PDT POP, which was designed using a condensation reaction between 6,6'-(1,4-phenylene)bis(1,3,5-triazine-2,4-diamine) (PDA-4NH2) and 4,4',4'',4'''-(pyrene-1,3,6,8-tetrayl)tetrabenzaldehyde (Py-Ph-4CHO) in the presence of dimethyl sulfoxide (DMSO) as a solvent. The obtained Py-PDT POP-600 with a high nitrogen content had a high surface area (up to 314 m2 g-1), high pore volume, and good thermal stability based on N2 adsorption/desorption data and a thermogravimetric analysis (TGA). Owing to the good surface area, the as-prepared Py-PDT POP-600 showed excellent performance in CO2 uptake (2.7 mmol g-1 at 298 K) and a high specific capacitance of 550 F g-1 at 0.5 A g-1 compared with the pristine Py-PDT POP (0.24 mmol g-1 and 28 F g-1).
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336MXenes, as highly electronegative and conductive two-dimensional nanomaterials, are extensively studied for their use in sensors and flexible electronics. In this study, near-field electrospinning was used to prepare a new poly(vinylidene difluoride) (PVDF)/Ag nanoparticle (AgNP)/MXene composite nanofiber film as a self-powered flexible human motion-sensing device. The composite film displayed highly piezoelectric properties with the presence of MXene. Scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy revealed that the intercalated MXene in the composite nanofibers was evenly spread out, which not only prevented the aggregation of MXene but also enabled the composite materials to form self-reduced AgNPs. The prepared PVDF/AgNP/MXene fibers displayed exceptional stability and excellent output performance, enabling their use for energy harvesting and powering light-emitting diodes. The doping of MXene/AgNPs increased the electrical conductivity of the PVDF material, improved its piezoelectric properties, and enhanced the piezoelectric constant of PVDF piezoelectric fibers, thereby allowing the production of flexible, sustainable, wearable, and self-powered electrical devices.
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Breath monitoring and pulmonary function analysis have been the prime focus of wearable smart sensors owing to the COVID-19 outbreak. Currently used lung function meters in hospitals are prone to spread the virus and can result in the transmission of the disease. Herein, we have reported the first-ever wearable patch-type strain sensor for enabling real-time lung function measurements (such as forced volume capacity (FVC) and forced expiratory volume (FEV) along with breath monitoring), which can avoid the spread of the virus. The noninvasive and highly sensitive strain sensor utilizes the synergistic effect of two-dimensional (2D) silver flakes (AgFs) and one-dimensional (1D) silver nanowires (AgNWs), where AgFs create multiple electron transmission paths and AgNWs generate percolation networks in the nanocomposite. The nanocomposite-based strain sensor possesses a high optimized conductivity of 7721 Sm-1 (and a maximum conductivity of 83,836 Sm-1), excellent stretchability (>1000%), and ultrasensitivity (GFs of 35 and 87 when stretched 0-20 and 20-50%, respectively), thus enabling reliable detection of small strains produced by the body during breathing and other motions. The sensor patching site was optimized to accurately discriminate between normal breathing, quick breathing, and deep breathing and analyze numerous pulmonary functions, including the respiratory rate, peak flow, FVC, and FEV. Finally, the observed measurements for different pulmonary functions were compared with a commercial peak flow meter and a spirometer, and a high correlation was observed, which highlights the practical feasibility of continuous respiratory monitoring and pulmonary function analysis.
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COVID-19 , Nanocompostos , Nanofios , Humanos , Prata , PulmãoRESUMO
In this study, polyethylene glycol (PEG) and polyurethane (PU)-based shape-stabilized copolymer nanocomposites were synthesized and utilized for developing low-cost and flexible temperature sensors. PU was utilized as a flexible structural material for loading a thermosensitive phase change PEG polymer by means of physical mixing and chemical crosslinking. Furthermore, the introduction of multi-walled carbon nanotubes (MWCNT) as a conductive filler in the PEG-PU copolymer resulted in a nanocomposite with thermoresistive properties. MWCNT loading concentrations from 2 wt.% to 10 wt.% were investigated, to attain the optimum conductivity of the nanocomposite. Additionally, the effect of MWCNT loading concentration on the thermosensitive behavior of the nanocomposite was analyzed in the temperature range 25 °C to 50 °C. The thermosensitive properties of the physically mixed and crosslinked polymeric nanocomposites were compared by spin coating the respective nanocomposites on screen printed interdigitated (IDT) electrodes, to fabricate the temperature sensor. The chemically crosslinked MWCNT-PEG-PU polymeric nanocomposite showed an improved thermosensitive behavior in the range 25 °C to 50 °C, compared to the physically mixed nanocomposite. The detailed structural, morphological, thermal, and phase transition properties of the nanocomposites were investigated using XRD, FTIR, and DSC analysis. XRD and FTIR were used to analyze the crystallinity and PEG-PU bonding of the copolymer nanocomposite, respectively; while the dual phase (solid-liquid) transition of PEG was analyzed using DSC. The proposed nanocomposite-based flexible temperature sensor demonstrated excellent sensitivity, reliability and shows promise for a wide range of bio-robotic and healthcare applications.
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Owing to their excellent sensitivity, stretchability, flexibility and conductivity, polymeric nanocomposites with conductive fillers have shown promise for a wide range of applications in bioelectronics and wearable devices. Herein, we report on the development of a flexible and biocompatible polymeric nanocomposite comprising ultra-long Ag-Au core-sheath nanowires (Au@AgNWs) dispersed in elastomeric media to fabricate a high-resolution wearable temperature sensor. Ultra-long AgNWs with an aspect ratio of about 1500 were synthesized using a Ca2+ ion-mediated facile one-pot polyol process. To enhance the biocompatibility and anti-oxidative property of the AgNWs, a 10-20 nm gold (Au) layer was conformably deposited without affecting the original nanowire morphology. The core-sheath structure of Au@AgNWs was characterized using HRTEM and EDS elemental mapping while the biocompatibility and anti-oxidative properties were tested using hydrogen peroxide (H2O2) etching in solution phase. Finally, the fabricated nanowires were used to prepare the Au@AgNW-poly-ethylene glycol (PEG)-polyurethane (PU)-based nanocomposite ink which can be printed on interdigitated electrodes to fabricate a thermoresistive temperature sensor with negative temperature coefficient (NTC) of resistance and quick response time (<100 s). The Au@AgNW-PEG-PU nanocomposite was characterized in detail and a novel temperature sensing mechanism based on controlling the internanowire distance of the PEG coated Au@AgNWs percolation by means of capillarity force among the nanowires as a result of the glass transition temperature of thermosensitive PEG was demonstrated. The proposed printable temperature sensor is flexible and biocompatible and shows promise for a range of wearable applications.
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Nanocompostos , Nanofios , Peróxido de Hidrogênio , Prata , TemperaturaRESUMO
Creatinine biosensing is a rapidly developing field owing to the clinical relevance of creatinine as a vital biomarker for several diseases associated with renal, thyroidal, and muscular dysfunctions. Over the years, we have observed numerous creatinine biosensing strategies, including the most widely studied enzymatic creatinine biosensors. Though the enzymatic approach provides excellent selectivity and reliability, it has certain drawbacks, which include high fabrication cost and poor storage stability (that is inherent to every enzyme-based biosensors). This has led to the development of non-enzymatic creatinine biosensors, of which electrochemical sensors are the most promising for point-of-care applications. However, only a limited number of studies have been conducted and there is a lack of reviews addressing the recent advances in this research area. Herein, we present for the first time, a review with a prime focus on the various strategies implemented in non-enzymatic electrochemical creatinine biosensing. We aim to offer a comprehensive context on the achievements and limitations of currently available non-enzymatic electrochemical creatinine biosensors and address the underlying factors pertaining to the interplay of modification/fabrication techniques with the sensitivity, selectivity, interferences, and long-term storage stability of the biosensor. We hope that this work shall prove to be seminal in the conception and advancement of future non-enzymatic electrochemical creatinine biosensors.
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Técnicas Biossensoriais , Creatinina , Sistemas Automatizados de Assistência Junto ao Leito , Reprodutibilidade dos TestesRESUMO
Piezoresistive tactile sensors made using nanocomposite polymeric materials have been shown to possess good flexibility, electrical performance, and sensitivity. However, the sensing performance, especially in the low-pressure range, can be significantly improved by enabling uniform dispersion of the filler material and utilization of effective structural designs that improve the tactile sensing performance. In this study, a novel flexible piezoresistive tactile sensor with a grid-type microstructure was fabricated using polymer composites comprising multi-walled carbon nanotubes (MWCNTs) as the conductive filler and polydimethylsiloxane (PDMS) as the polymeric matrix. The research focused on improving the tactile sensor performance by enabling uniform dispersion of filler material and optimizing sensor design and structure. The doping weight ratio of MWCNTs in PDMS varied from 1 wt.% to 10 wt.% using the same grid structure-sensing layer (line width, line spacing, and thickness of 1 mm). The sensor with a 7 wt.% doping ratio had the most stable performance, with an observed sensitivity of 6.821 kPa-1 in the lower pressure range of 10-20 kPa and 0.029 kPa-1 in the saturation range of 30-200 kPa. Furthermore, the dimensions of the grid structure were optimized and the relationship between grid structure, sensitivity, and sensing range was correlated. The equation between pressure and resistance output was derived to validate the principle of piezoresistance. For the grid structure, dimensions with line width, line spacing, and thickness of 1, 1, and 0.5 mm were shown to have the most stable and improved response. The observed sensitivity was 0.2704 kPa-1 in the lower pressure range of 50-130 kPa and 0.0968 kPa-1 in the saturation range of 140-200 kPa. The piezoresistive response, which was mainly related to the quantum tunneling effect, can be optimized based on the dopant concentration and the grid microstructure. Furthermore, the tactile sensor showed a repeatable response, and the accuracy was not affected by temperature changes in the range of 10 to 40 °C and humidity variations from 50 to 80%. The maximum error fluctuation was about 5.6% with a response delay time of about 1.6 ms when cyclic loading tests were performed under a normal force of 1 N for 10,200 cycles. Consequently, the proposed tactile sensor shows practical feasibility for a wide range of wearable technologies and robotic applications such as touch detection and grasping.
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One-dimensional metal nanowires, with novel functionalities like electrical conductivity, optical transparency and high mechanical stiffness, have attracted widespread interest for use in applications such as transparent electrodes in optoelectronic devices and active components in nanoelectronics and nanophotonics. In particular, silver nanowires (AgNWs) have been widely researched owing to the superlative thermal and electrical conductivity of bulk silver. Herein, we present a detailed review of the synthesis of AgNWs and their utilization in fabricating improved transparent conducting electrodes (TCE). We discuss a range of AgNW synthesis protocols, including template assisted and wet chemical techniques, and their ability to control the morphology of the synthesized nanowires. Furthermore, the use of scalable and cost-effective solution deposition methods to fabricate AgNW based TCE, along with the numerous treatments used for enhancing their optoelectronic properties, are also discussed.
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Galectin-1 protein has been recently recognized as a valuable urinary biomarker for the diagnosis and prognosis of bladder cancer. Herein, we present a sensitive and specific impedimetric immunosensor for the quantitative and label free detection of Galectin-1 protein in clinical urine samples. The immunosensor consists of nine gold interdigitated microelectrodes (3 × 3 array), which can simultaneously monitor multiple immunoreactions by analyzing the normalized impedance variations at each microelectrode during immunosensing. To obtain enhanced sensitivities, we have utilized Galectin-1/Al2O3 nanoprobes (Galectin-1 antibody conjugated to alumina nanoparticles) that can be selectively trapped on the microelectrode surface using positive dielectrophoresis (p-DEP). Preliminary studies highlight the feasibility of the proposed immunosensor for Gal -1 detection in T24 cell lysate spiked phosphate buffer saline and artificial urine samples with a limit of detection that is estimated to be in the pg/ml range. To verify its practical feasibility, we have tested the immunosensor for Galectin-1 detection in clinical urine samples obtained from normal patients and those diagnosed with bladder cancer. Analysis of the clinical tests shows that the median normalized impedance variation during immunosensing for 22 cancer patients and 26 normal patients is 27% and 10%, respectively, with an identified cutoff point of 19.5% above which the sensitivity and specificity of bladder cancer detection was 82.1% and 80.8%, respectively. Based on these results, the proposed immunosensor shows promise for bladder cancer diagnosis and prognosis in a point of care format, thus enabling improved public health monitoring.
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Técnicas Biossensoriais/instrumentação , Imunoensaio/instrumentação , Neoplasias da Bexiga Urinária/diagnóstico , Neoplasias da Bexiga Urinária/urina , Estudos de Casos e Controles , Linhagem Celular Tumoral , Impedância Elétrica , Galectina 1/urina , Humanos , Neoplasias da Bexiga Urinária/patologiaRESUMO
The presence of small amounts of human serum albumin (HSA) in urine or microalbuminuria (30-300 µg/mL) is a valuable clinical biomarker for the early detection of chronic kidney disease (CKD). Herein, we report on the development of an inexpensive and disposable immunosensor for the sensitive, specific, and label-free detection of HSA using electrochemical impedance spectroscopy (EIS). We have utilized a simple one-step screen-printing protocol to fabricate the carbon-based three-electrode system on flexible plastic substrates. To enable efficient antibody immobilization and improved sensitivity, the carbon working electrode was sequentially modified with electropolymerized polyaniline (PANI) and electrodeposited gold nanocrystals (AuNCs). The PANI matrix serves as an interconnected nanostructured scaffold for homogeneous distribution of AuNCs and the resulting PANI/AuNCs nanocomposite synergically improved the immunosensor response. The PANI/AuNCs-modified working electrode surface was characterized using scanning electron microscopy (SEM) and the electrochemical response at each step was analyzed using EIS in a ferri/ferrocyanide redox probe solution. The normalized impedance variation during immunosensing increased linearly with HSA concentration in the range of 3-300 µg/mL and a highly repeatable response was observed for each concentration. Furthermore, the immunosensor displayed high specificity when tested using spiked sample solutions containing different concentrations of actin protein and J82 cell lysate (a complex fluid containing a multitude of interfering proteins). Consequently, these experimental results confirm the feasibility of the proposed immunosensor for early diagnosis and prognosis of CKD at the point of care.
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Compostos de Anilina/química , Técnicas Biossensoriais/métodos , Ouro/química , Nanocompostos/química , Insuficiência Renal Crônica/diagnóstico , Carbono/química , Linhagem Celular Tumoral , Espectroscopia Dielétrica , Eletrodos , Humanos , Nanopartículas Metálicas/química , Albumina Sérica Humana/análise , Propriedades de SuperfícieRESUMO
Several wearable devices have already been commercialized and are likely to open up a new life pattern for consumers. However, the limited energy capacity and lifetime have made batteries the bottleneck in wearable technology. Thus, there have been growing efforts in the area of self-powered wearables that harvest ambient mechanical energy directly from surroundings. Herein, we demonstrate a woven triboelectric nanogenerator (WTENG) utilizing electrospun Polyvinylidene fluoride (PVDF) nanofibers and commercial nylon cloth to effectively harvest mechanical energy from human motion. The PVDF nanofibers were fabricated using a highly scalable multi-nozzle far-field centrifugal electrospinning protocol. We have also doped the PVDF nanofibers with small amounts of multi-walled carbon nanotubes (MWCNT) to improve their triboelectric performance by facilitating the growth of crystalline ß-phase with a high net dipole moment that results in enhanced surface charge density during contact electrification. The electrical output of the WTENG was characterized under a range of applied forces and frequencies. The WTENG can be triggered by various free-standing triboelectric layers and reaches a high output voltage and current of about 14 V and 0.7 µA, respectively, for the size dimensions 6 × 6 cm. To demonstrate the potential applications and feasibility for harvesting energy from human motion, we have integrated the WTENG into human clothing and as a floor mat (or potential energy generating shoe). The proposed triboelectric nanogenerator (TENG) shows promise for a range of power generation applications and self-powered wearable devices.
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In this study, we have fabricated a simple disposable electrochemical immunosensor for the point of care testing of microalbuminuria, a well-known clinical biomarker for the onset of chronic kidney disease. The immunosensor is fabricated by screen-printing carbon interdigitated microelectrodes on a flexible plastic substrate and utilizes electrochemical impedance spectroscopy to enable direct and label free immunosensing by analyzing interfacial changes on the electrode surface. To improve conductivity and biocompatibility of the screen-printed electrodes, we have modified it with gold nanoparticles, which are electrodeposited using linear sweep voltammetry. To enable efficient immobilization of HSA antibodies, we have developed novel PS/Ag/ab-HSA nanoprobes (polystyrene nanoparticle core with silver nanoshells covalently conjugated to HSA antibodies), and these nanoprobes are trapped on the electrode surface using dielectrophoresis. Each immunosensor has two sensing sites corresponding to test and control to improve specificity by performing differential analysis. Immunosensing results show that the normalized impedance response is linearly dependent on albumin concentration in the clinically relevant range with good repeatability. We have also developed a portable impedance readout module that can analyze the data obtained from the immunosensor and transmit it wirelessly for cloud computing. Consequently, the developed immunosensing platform can be extended to the detection of a range of immunoreactions and shows promise for point of diagnosis and public healthcare monitoring.
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Técnicas Biossensoriais , Imunoensaio , Imunoconjugados/química , Nanopartículas/química , Anticorpos Imobilizados/química , Anticorpos Imobilizados/imunologia , Carbono/química , Ouro/química , Humanos , Limite de Detecção , Nanopartículas Metálicas/química , Nanoconchas/química , Poliestirenos/químicaRESUMO
In this study, we have developed a novel paper based immunoassay for the quantitative detection of immunoreactions using electrochemical impedance spectroscopy. Paper provides an attractive platform for fabrication of simple, low cost, and portable diagnostic devices as it allows passive liquid transport, is biocompatible, and has tunable properties such as hydrophilicity, flexibility, permeability, and reactivity. We have used screen-printing to fabricate interdigitated electrodes (finger width and gap of 200 µm) on the paper substrate, while UV-lithography enables patterning of the paper into hydrophobic/hydrophilic regions. As a proof of concept, we have used this immunosensor to detect the immune response of Human Serum Albumin (HSA) antibody-antigen complex formation. To enable efficient immobilization of HSA antibodies, we have utilized dielectrophoresis to trap microprobes (MPs) on the electrode surface. The microprobes consist of an alumina nanoparticle core with a well-adhered polyaniline outer coating to which the HSA antibodies are conjugated in an oriented manner via covalent chemistry. The efficacy of the impedance-based immunosensor is compared when MPs are immobilized specifically on the electrode surface using dielectrophoresis (DEP) as opposed to being dropped and immobilized via physical absorption on the entire sensing area. Results show that a more reproducible and sensitive response is observed when DEP is utilized to trap the microprobes. Furthermore, the normalized impedance variation during immunosensing shows a linear dependence on the concentration of HSA with an observed limit of detection of 50 µg/ml, which is lower than conventionally used paper based urine dipsticks used for urinary protein detection. Thus, we have developed a low cost paper based immunoassay platform that can be used for the quantitative point of care detection of a wide range of immunoreactions.
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An ultrasensitive and real-time impedance based immunosensor has been fabricated for the quantitative detection of Galectin-1 (Gal-1) protein, a biomarker for the onset of multiple oncological conditions, especially bladder cancer. The chip consists of a gold annular interdigitated microelectrode array (3×3 format with a sensing area of 200µm) patterned using standard microfabrication processes, with the ability to electrically address each electrode individually. To improve sensitivity and immobilization efficiency, we have utilized nanoprobes (Gal-1 antibodies conjugated to alumina nanoparticles through silane modification) that are trapped on the microelectrode surface using programmable dielectrophoretic manipulations. The limit of detection of the immunosensor for Gal-1 protein is 0.0078mg/ml of T24 (Grade III) cell lysate in phosphate buffered saline, artificial urine and human urine samples. The normalized impedance variations show a linear dependence on the concentration of cell lysate present while specificity is demonstrated by comparing the immunosensor response for two different grades of bladder cancer cell lysates. We have also designed a portable impedance analyzing device to connect the immunosensor for regular checkup in point of care testing with the ability to transfer data over the internet using a personal computer. We believe that this diagnostic system would allow for improved public health monitoring and aid in early cancer diagnosis.
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Técnicas Biossensoriais/instrumentação , Galectina 1/urina , Testes Imediatos , Neoplasias da Bexiga Urinária/urina , Óxido de Alumínio/química , Anticorpos Imobilizados/química , Linhagem Celular Tumoral , Impedância Elétrica , Desenho de Equipamento , Humanos , Imunoensaio/instrumentação , Limite de Detecção , Microeletrodos , Nanopartículas/química , Neoplasias da Bexiga Urinária/diagnósticoRESUMO
A multiplexed immunosensor has been developed for the detection of specific biomarkers Galectin-1 (Gal-1) and Lactate Dehydrogenase B (LDH-B) present in different grades of bladder cancer cell lysates. In order to immobilize nanoprobes with different antibodies on a single chip we employed three-step programmable dielectrophoretic manipulations for focusing, guiding and trapping to enhance the fluorescent response and reduce the interference between the two antibody arrays. The chip consisted of a patterned indium tin oxide (ITO) electrode for sensing and a middle fish bone shaped gold electrode for focusing and guiding. Using ITO electrodes for the sensing area can effectively eliminate the background noise of fluorescence response as compared to metal electrodes. It was also observed that the three step manipulation increased fluorescence response after immunosensing by about 4.6 times as compared to utilizing DEP for just trapping the nanoprobes. Two different-grade bladder cancer cell lysates (grade I: RT4 and grade III: T24) were individually analyzed for detecting the protein expression levels of Gal-1 and LDH-B. The fluorescence intensity observed for Gal-1 is higher than that of LDH-B in the T24 cell lysate; however the response observed in RT4 is higher for LDH-B as compared to Gal-1. Thus we can effectively identify the different grades of bladder cancer cells. In addition, the platform for DEP manipulation developed in this study can enable real time detection of multiple analytes on a single chip and provide more practical benefits for clinical diagnosis.