Performance Optimization of Wearable Printed Human Body Temperature Sensor Based on Silver Interdigitated Electrode and Carbon-Sensing Film.

The human body's temperature is one of the most important vital markers due to its ability to detect various diseases early. Accurate measurement of this parameter has received considerable interest in the healthcare sector. We present a novel study on the optimization of a temperature sensor based on silver interdigitated electrodes (IDEs) and carbon-sensing film. The sensor was developed on a flexible Kapton thin film first by inkjet printing the silver IDEs, followed by screen printing a sensing film made of carbon black. The IDE finger spacing and width of the carbon film were both optimized, which considerably improved the sensor's sensitivity throughout a wide temperature range that fully covers the temperature of human skin. The optimized sensor demonstrated an acceptable temperature coefficient of resistance (TCR) of 3.93 × 10-3 °C-1 for temperature sensing between 25 °C and 50 °C. The proposed sensor was tested on the human body to measure the temperature of various body parts, such as the forehead, neck, and palm. The sensor showed a consistent and reproducible temperature reading with a quick response and recovery time, exhibiting adequate capability to sense skin temperatures. This wearable sensor has the potential to be employed in a variety of applications, such as soft robotics, epidermal electronics, and soft human-machine interfaces.

Bench-Top Fabrication of an All-PDMS Microfluidic Electrochemical Cell Sensor Integrating Micro/Nanostructured Electrodes.

In recent years, efforts in the development of lab-on-a-chip (LoC) devices for point-of-care (PoC) applications have increased to bring affordable, portable, and sensitive diagnostics to the patients' bedside. To reach this goal, research has shifted from using traditional microfabrication methods to more versatile, rapid, and low-cost options. This work focuses on the benchtop fabrication of a highly sensitive, fully transparent, and flexible poly (dimethylsiloxane) (PDMS) microfluidic (µF) electrochemical cell sensor. The µF device encapsulates 3D structured gold and platinum electrodes, fabricated using a shape-memory polymer shrinking method, which are used to set up an on-chip electrochemical cell. The PDMS to PDMS-structured electrode bonding protocol to fabricate the µF chip was optimized and found to have sufficient bond strength to withstand up to 100 mL/min flow rates. The sensing capabilities of the on-chip electrochemical cell were demonstrated by using cyclic voltammetry to monitor the adhesion of murine 3T3 fibroblasts in the presence of a redox reporter. The charge transfer across the working electrode was reduced upon cell adhesion, which was used as the detection mechanism, and allowed the detection of as few as 24 cells. The effective utilization of simple and low cost bench-top fabrication methods could accelerate the prototyping and development of LoC technologies and bring PoC diagnostics and personalized medicine to the patients' bedside.

Screen-printed, biocompatible and ultrasensitive sensor for real-time reactive oxygen species detection in human sweat.

Excessive or burst generation of reactive oxygen species (ROS) can induce oxidative stress, precipitating a range of critical illnesses, including cancers, Parkinson's disease and Ischemia-reperfusion injury. Conventional biological assays for ROS, involving discrete steps of capturing, labelling, and spectrometric detection, are complex and time-intensive. Moreover, their accuracy is substantially compromised by the short lifespan (microseconds to milliseconds) of ROS. Consequently, there is a pressing need for a rapid and efficient method that enables real-time detection. In this study, we have developed a printable, flexible ROS sensor based on a robust nanoenzyme composite by direct deposition of the paste onto a flexible polyethylene terephthalate (PET) substrate. This device demonstrated the fast and real-time responses to the hydrogen peroxide (mimetic agent) in the laboratory and to total ROS in sweat of an individual, exhibiting an outstanding current response to hydrogen peroxide across a broad concentration range of 0.01-10 mM, with a limit of detection (LOD) of 1.85 µM. The device's sensitivity to hydrogen peroxide (136.59 µA mM-1 cm-2), was found to be 1.5 to 10 times higher than that of sensors previously reported. Moreover, the IFRS device successfully identified instantaneous ROS levels in the sweat of adult males in vitro, with amperometric response increased 8 times after half an hour strenuous exercise, thereby exhibiting excellent selectivity, remarkable stability, and confirmed high biosafety. Overall, the IFRS provides a viable and practical solution for simple, expedited, and real-time ROS detection in the near future.

Ultra-thin 2D bimetallic MOF nanosheets for highly sensitive and stable detection of glucose in sweat for dancer.

The measurement of glucose concentration in sweat is expected to replace the existing blood glucose detection, which realize the effective way of non-invasive monitoring of human glucose concentration in dancing. High precision glucose detection can be achieved by adjusting the electrode material of the sensor. Thus, in this work, the bimetallic organic frameworks (bi-MOFs) materials containing Mn and Ni ions (NiMn-MOF) with ultrathin nanosheets have been exquisitely designed. The ultrathin nanosheet and heterogeneous metal ions in the structure optimize the electronic structure, which improves the electrical conductivity of MOFs. The success of the preparation strategy leads the good electrocatalytic performance of NiMn-MOF for glucose detection. Detailedly, NiMn-MOF shows high sensitivity of 1576 µA mM-1 cm-2 in the linear range from 0 to 0.205 mM and the wide linear region of 0.255-2.655 mM and 3.655-5.655 mM were also observed. In addition, the high repeatability, reproductivity, long-term stability and ultra-low limited of detection (LOD, 0.28 µM, S/N = 3) provide foundation for the practical sensor application of this NiMn-MOF nanosheets. Remarkably, as designed NiMn-MOF sensor can accurately measure glucose in sweat showing great potential in the field of wearable glucose monitoring during dancing.

Label free flexible electrochemical DNA biosensor for selective detection of Shigella flexneri in real food samples.

An effective tool for early-stage selective detection of the foodborne bacterial pathogen Shigella flexneri (S. flexneri) is essential for diagnosing infectious diseases and controlling outbreaks. Here, a label-free electrochemical DNA biosensor for monitoring S. flexneri is developed. To fabricate the biosensor, detection probe (capture probe) is immobilized on the surface of poly melamine (P-Mel) and poly glutamic acid (PGA), and disuccinimidyl suberate (DSS) functionalized flexible indium tin oxide (ITO) electrode. Anthraquinone-2-sulfonic acid monohydrate sodium salt (AQMS) is used as a signal indicator for the detection of S. flexneri. The proposed DNA biosensor exhibits a wide dynamic range with concentration of the targets ranging from 1 × 10-6 to 1 × 10-21 molL-1 with a limit of detection (LOD) of 7.4 × 10-22 molL-1 in the complementary linear target of S. flexneri, and a detection range of 8 × 1010-80 cells/ml with a LOD of 10 cells/ml in real S. flexneri sample. The proposed flexible biosensor provides high specificity for the detection of S. flexneri compared to other target signals such as discrete base mismatches and different bacterial species. The developed biosensor displayed excellent recoveries in detecting S. flexneri in spiked food samples. Therefore, the proposed biosensor can serve as a model methodology for the detection of other pathogens in a broad span of industries.

Corrigendum: Flexible biosensors based on colorimetry, fluorescence, and electrochemistry for point-of-care testing.

[This corrects the article DOI: 10.3389/fbioe.2021.753692.].

Graphene Microelectrodes for Real-Time Impedance Spectroscopy of Neural Cells.

Understanding the changes in the electrochemical properties of neural cells upon exposure to stress factors imparts vital information about the conditions prior to their death. This study presents a graphene-based biosensor for real-time monitoring of N27 rat dopaminergic neural cells which characterizes cell adhesion and cytotoxicity factors through impedance spectroscopy. The aim was to monitor the growth of the entire cell network via a nonmetallic flexible electrode array. Therefore, a water-based graphene solution was formulized as a conductive ink, 3D-printed into a flexible substrate through an electrohydrodynamic approach, resulting in electrodes with a conductivity of 6750 s/m. The presented high-throughput method enabled microscale monitoring of the entire cell network via the design of PDMS-based growth channels. The electrical resistance of the cell network was measured continuously along with their network density, constituting a mean density of 1890 cell/mm2 at full cell confluency. The results demonstrate the applicability of the impedance-based sensing of the cell network for rapid screening of the cytotoxic elements, and the real-time effect of UV exposure on dopaminergic neural cells was reported as an immediate application of the device.

Flexible triphase enzyme electrode based on hydrophobic porous PVDF membrane for high-performance bioassays.

Flexible bioassays based on oxidase-catalyzed and electrocatalytic cascade reactions have been widely reported. However, the fluctuant oxygen level and high anodic potential restricts the detection accuracy. To overcome these challenges, we report here a flexible triphase enzyme electrode by assembling an oxidase enzyme layer and Pt electrocatalysts onto a carbon nanotube film/porous polyvinylidene fluoride hydrophobic substrate. Such a flexible enzyme electrode has an air-liquid-solid triphase reaction zone where oxygen level is air phase dependent (constant and sufficient high), which stabilized the oxidase kinetics and enabled the cathodic measurement of enzymatic product H2O2 with minimum interferents caused from oxygen level fluctuation and many oxidizable species in analyte solution. Furthermore, the flexible triphase enzyme electrode exhibited good mechanical stability even after being bent over 600 times and an excellent air permeability, which are crucial to wearable devices that require long-term skin contact.

Towards a flexible electrochemical biosensor fabricated from biocompatible Bombyx mori silk.

In modern days, there is an increasing relevance of and demand for flexible and biocompatible sensors for in-vivo and epidermal applications. One promising strategy is the implementation of biological (natural) polymers, which offer new opportunities for flexible biosensor devices due to their high biocompatibility and adjustable biodegradability. As a proof-of-concept experiment, a biosensor was fabricated by combining thin- (for Pt working- and counter electrode) and thick-film (for Ag/AgCl quasi-reference electrode) technologies: The biosensor consists of a fully bio-based and biodegradable fibroin substrate derived from silk fibroin of the silkworm Bombyx mori combined with immobilized enzyme glucose oxidase. The flexible glucose biosensor is encapsulated by a biocompatible silicon rubber which is certificated for a safe use onto human skin. Characterization of the sensor set-up is exemplarily demonstrated by glucose measurements in buffer and Ringer's solution, while the stability of the quasi-reference electrode has been investigated versus a commercial Ag/AgCl reference electrode. Repeated bending studies validated the mechanical properties of the electrode structures. The cross-sensitivity of the biosensor against ascorbic acid, noradrenaline and adrenaline was investigated, too. Additionally, biocompatibility and degradation tests of the silk fibroin with and without thin-film platinum electrodes were carried out.

Flexible Biosensors Based on Colorimetry, Fluorescence, and Electrochemistry for Point-of-Care Testing.

With the outbreak and pandemic of COVID-19, point-of-care testing (POCT) systems have been attracted much attention due to their significant advantages of small batches of samples, user-friendliness, easy-to-use and simple detection. Among them, flexible biosensors show practical significance as their outstanding properties in terms of flexibility, portability, and high efficiency, which provide great convenience for users. To construct highly functional flexible biosensors, abundant kinds of polymers substrates have been modified with sufficient properties to address certain needs. Paper-based biosensors gain considerable attention as well, owing to their foldability, lightweight and adaptability. The other important flexible biosensor employs textiles as substrate materials, which has a promising prospect in the area of intelligent wearable devices. In this feature article, we performed a comprehensive review about the applications of flexible biosensors based on the classification of substrate materials (polymers, paper and textiles), and illustrated the strategies to design effective and artificial sensing platforms, including colorimetry, fluorescence, and electrochemistry. It is demonstrated that flexible biosensors play a prominent role in medical diagnosis, prognosis, and healthcare.

Flexible sensor based on conducting polymer and gold nanoparticles for electrochemical screening of HPV families in cervical specimens.

Considering the low sensitivity of cytological exams and high costs of the molecular methods, the development of diagnostic tests for effective diagnosis of HPV infections is a priority. In this work, biosensor composed of polypyrrole (PPy) films and gold nanoparticles (AuNPs) was obtained for specific detection of HPV genotypes. The biosensor was developed by using flexible electrodes based on polyethylene terephthalate (PET) strips coated with indium tin oxide (ITO). Polymeric films and AuNPs were obtained by electrosynthesis. Oligonucleotides sequences modified with functional amino groups were designed to recognize HPV gene families strictly. The modified oligonucleotides were chemically immobilized on the nanostructured platform. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used for the analysis of the electrode modification and monitoring of molecular hybridization. Electrochemical changes were observed after exposure of the biosensors to plasmid samples and cervical specimens. The biosensor based on the BSH16 probe showed a linear concentration range for target HPV16 gene detection of 100 pg µL-1 to 1 fg µL-1. A limit of detection (LOD) of 0.89 pg µL-1 and limit of quantification (LOQ) of 2.70 pg µL-1 were obtained, with a regression coefficient of 0.98. Screening tests on cervical specimens were performed to evaluate the sensibility and specificity for HPV and its viral family. The expression of a biomarker for tumorigenesis (p53 gene) was also monitored. In this work, a flexible system has been successfully developed for label-free detection of HPV families and p53 gene monitoring with high specificity, selectivity, and sensitivity.

Novel Cellulose Fibre-Based Flexible Plasmonic Membrane for Point-of-Care SERS Biomarker Detection in Chronic Wound Healing.

BACKGROUND: Wound management is stretching the limits of health systems globally, challenging clinicians to evaluate the effectiveness of their treatments and deliver appropriate care to their patients. Visual inspection and manual measurement of wound size are subjective, often inaccurate and inconsistent. Growth factors, such as pro-inflammatory cytokines and proteases, play important roles in cutaneous wound healing. However, little is known about the point-of-care monitoring of the changes in such markers during the healing process. Here, we explore the capability of surface-enhanced Raman spectroscopy (SERS) as a viable point-of-care platform to monitor the changes of these surrogate indicators of healing status in chronic wounds. METHODS: We developed a biofunctionalized flexible, cost-effective, scalable and easy-to-fabricate plasmonic SERS substrate using cellulose fibre (CF), which is used for sensing of wound markers based on a modified immunoassay method. RESULTS: We evaluated and selected the reliable silver nano-island thickness that will be sputtered onto the CF-based substrate for the highest SERS enhancement. Using this biofunctionalized SERS substrate, we detected varying concentrations of MMP-9 (10-5000 ng/mL) and TNF-α (5-100 ng/mL) proteins to model the wound exudates. This SERS detection method demonstrates a linear response within biologically relevant concentrations, ranging from 10 to 500 ng/mL for MMP-9 and 5 to 25 ng/mL for TNF-α for these surrogate indicators. CONCLUSION: Our SERS sensing platform achieved detection limits in the µM to sub-nM range and displayed high sensitivity and selectivity. This could result in a cheap, point-of-care device that provides a non-invasive measure of cutaneous wound healing in real time. We envision that these flexible substrates after activation may be incorporated into wound dressings in future for routine monitoring of wound healing status.

A wearable battery-free wireless and skin-interfaced microfluidics integrated electrochemical sensing patch for on-site biomarkers monitoring in human perspiration.

In this study, an ultra-high sensitive, flexible, wireless, battery-free, and fully integrated (no external analysis equipment) electrochemical sensing patch system, including a microfluidic-sweat collecting unit, was newly developed for the on-site monitoring of the [K+] concentration in human sweat. Multiwalled carbon nanotube (MWCNT) and MXene-Ti3C2TX based hybrid multi-dimensional networks were applied to obtain a high surface activation area and faster charge transfer rate, strongly adsorbing the valinomycin membrane to protect the ionophore for effective transshipment and immobilization of the [K+]. Furthermore, the controllable porosity of carbon-based materials can accelerate the kinetic process of ion diffusion. This hybrid nanonetwork structure effectively enhanced electrochemical stability and sensitivity, addressing the noise and signal drifting problems experienced with low concentration detection. The fabricated sensor exhibited a high ion concentration sensitivity of 63 mV/dec with excellent selectivity, amplified to 173 mV/dec with the integrated amplification system. The Near Field Communication (NFC) is used to transmit measurements to a smartphone wirelessly. A microfluidic channel was integrated with the electrochemical sensor patch to efficiently collect sweat on the human skin surface and mitigate the sensor surface contamination problem. Furthermore, the developed sensing patch can also be applied to other biomarkers on-site detection after modifying the working electrode with the corresponding selective membranes.

Fabrication and Specific Functionalisation of Carbon Fibers for Advanced Flexible Biosensors.

This review aims at offering an up-to-date comprehensive summary of carbon fibers (CFs)-based composites, with the emphasis on smart assembly and purpose-driven specific functionalization for their critical applications associated with flexible sensors. We first give a brief introduction to CFs as a versatile building block for preparation of mutil-fountional materials and the current status of research studies on CFs. This is followed by addressing some crucial methods of preparation of CFs. We then summarize multiple possibilities of functionalising CFs, an evaluation of some key applications of CFs in the areas of flexible biosensors was also carried out.

Construction of flexible enzymatic electrode based on gradient hollow fiber membrane and multi-wall carbon tubes meshes.

In this study, we developed a convenient way to construct a flexible enzymatic electrode with excellent stability and electrochemical performance for implanted glucose monitoring. The electrode was constructed through the co-immobilization of the glucose oxidase micro-particles (GOD MPs) and multi-wall carbon nanotubes (CNT) on the inner surface of a gradient-structured hollow fiber membrane (GHM), where CNT improved the electron transport efficiency and GHM controlled the transfer of substances and interferences. GOD MPs showed higher stability under various operation conditions than the free enzymes due to the MnCO3 template method, which enabled the biosensor to remain relative sensitivity at >86% over 9 days. The GOD MPs biosensor also showed high selectivity, reproducibility, and linear sensing range from 0 mM to 24 mM (R2 = 0.9993) with a current sensitivity of 25 nA/mM. The combination of porous-structured membrane and the flexible CNT meshes ensures the electrical connections and sensing accuracy of the biosensor under the deformation status. In-vivo experiments showed reliable current responses to variations in blood glucose concentrations that were consistent with tail blood test results. This co-immobilization of enzyme micro-particles in the 3D porous structure method developed a bio-composite platform technology towards the applications in flexible sensing and implantable medical devices.

Tailoring the Meso-Structure of Gold Nanoparticles in Keratin-Based Activated Carbon Toward High-Performance Flexible Sensor.

Flexible biosensors with high accuracy and reliable operation in detecting pH and uric acid levels in body fluids are fabricated using well-engineered metal-doped porous carbon as electrode material. The gold nanoparticles@N-doped carbon in situ are prepared using wool keratin as both a novel carbon precursor and a stabilizer. The conducting electrode material is fabricated at 500 °C under customized parameters, which mimics A-B type (two different repeating units) polymeric material and displays excellent deprotonation performance (pH sensitivity). The obtained pH sensor exhibits high pH sensitivity of 57 mV/pH unit and insignificant relative standard deviation of 0.088%. Conversely, the composite carbon material with sp2 structure prepared at 700 °C is doped with nitrogen and gold nanoparticles, which exhibits good conductivity and electrocatalytic activity for uric acid oxidation. The uric acid sensor has linear response over a range of 1-150 µM and a limit of detection 0.1 µM. These results will provide new avenues where biological material will be the best start, which can be useful to target contradictory applications through molecular engineering at mesoscale.

Highly flexible and conductive poly (3, 4-ethylene dioxythiophene)-poly (styrene sulfonate) anchored 3-dimensional porous graphene network-based electrochemical biosensor for glucose and pH detection in human perspiration.

The patterned LIG flakes are generally not interconnected due to the line gap of the laser ray, leading to lower uniform conductivity and fragile graphene. Thus, the fabrication of a highly conductive and mechanically robust LIG-based biosensing platform remains challenging. In this study, the fabrication of a flexible electrochemical biosensor is reported based on poly (3, 4-ethylene dioxythiophene)-poly (styrene sulfonate) (PEDOT:PSS) modified 3-dimensional (3D) stable porous laser-induced graphene (LIG) for the detection of glucose and pH. PEDOT:PSS was spray-coated on the LIG to improve electrode robustness and deliver uniform electrical conductivity. The as-prepared PEDOT:PSS modified LIG (PP/LIG) was characterized using field-emission scanning electron microscopy (FESEM), x-ray photoelectron spectroscopy (XPS), Raman spectroscopy, and Fourier-transform infrared spectroscopy (FTIR). Platinum and palladium nanoparticles (Pt@Pd) were successfully electrodeposited on PP/LIG, markedly enhancing the electrocatalytic activity for glucose detection. The fabricated biosensor exhibited an excellent amperometric response to glucose with a wide linear range of 10 µM - 9.2 mM, a high sensitivity of 247.3 µAmM-1cm-2, and a low detection limit (LOD) of 3 µM, with high selectivity. In addition, the pH sensor was functionalized by the polyaniline (PANI) on PP/LIG, and it also exhibited excellent potentiometric response with a high sensitivity of 75.06 mV/pH in the linear range of pH 4 - 7. Ultimately, the feasibility of the biosensor was confirmed by the analysis of human perspiration collected during physical exercise. This approach validates the utility of the novel fabrication procedure, and the potential of the LIG-conductive polymer composite for biosensing applications.

Conformal, waterproof electronic decals for wireless monitoring of sweat and vaginal pH at the point-of-care.

While the monitoring of pH has demonstrated to be an effective technique to monitor an individual's health state, the design of wearable biosensors is subject to critical challenges, such as high fabrication costs, thermal drift, sensitivity to moisture, and the limited applicability for users with metal allergies. This work describes the low-cost fabrication of waterproof electronic decals (WPEDs): highly conformable disposable biosensors capable of monitoring sweat and vaginal pH. WPEDs contain a polyaniline/silver microflakes sensing layer optimized for accurate impedance-based pH quantification across the clinically relevant range of variation of most biofluids. WPEDs also contain a heating layer that serves to both stimulate sweating and prevent saturation of the sensing area, reducing the variability of the measurements. The conformability of WPEDs enables their simple and allergy-free attachment to skin, where they can monitor sweat pH, or to the surface of paper-based sample containers, for the pH-based diagnosis of bacterial vaginosis. WPEDs are mostly transparent, self-adhesive, breathable, flexible, moisture-insensitive, and able to maintain their accuracy under significant mechanical and thermal stresses. A cost-effective wearable and portable impedance analyzer wirelessly transmits pH data in real-time to the smartphone of the user, where a custom-developed App enables long term monitoring and telemedicine applications. Our results demonstrate the feasibility of using inexpensive single-use WPEDs and a reusable, wireless impedance analyzer to provide a wearable solution for the real-time monitoring of sweat pH and the accurate at-home diagnosis of bacterial vaginosis, improving the capabilities of current low-cost, point-of-care diagnostic tests.

Flexible HIV-1 Biosensor Based on the Au/MoS2 Nanoparticles/Au Nanolayer on the PET Substrate.

An electrochemical flexible biosensor composed of gold (Au), molybdenum disulfide nanoparticles (MoS2 NPs), and Au (Au/MoS2/Au nanolayer) on the polyethylene terephthalate (PET) substrate is developed to detect envelope glycoprotein GP120 (gp120), the surface protein of HIV-1. To fabricate the nanolayer on the PET substrate, Au is sputter coated on the flexible PET substrate and MoS2 NPs are spin coated on Au, which is sputter coated once again with Au. The gp120 antibody is then immobilized on this flexible electrode through cysteamine (Cys) modified on the surface of the Au/MoS2/Au nanolayer. Fabrication of the biosensor is verified by atomic force microscopy, scanning electron microscopy, and cyclic voltammetry. A flexibility test is done using a micro-fatigue tester. Detection of the gp120 is measured by square wave voltammetry. The results indicate that the prepared biosensor detects 0.1 pg/mL of gp120, which is comparable with previously reported gp120 biosensors prepared even without flexibility. Therefore, the proposed biosensor supports the development of a nanomaterial-based flexible sensing platform for highly sensitive biosensors with flexibility for wearable device application.

Flexible Biosensors for the Impedimetric Detection of Protein Targets Using Silk-Conductive Polymer Biocomposites.

To expand the applications of flexible biosensors in point-of-care healthcare applications beyond monitoring of biophysical parameters, it is important to devise strategies for the detection of various proteins and biomarkers. Here, we demonstrate a flexible, fully organic, biodegradable, label-free impedimetric biosensor for the critical biomarker, vascular endothelial growth factor (VEGF). This biosensor was constructed by photolithographically patterning a conducting ink consisting of a photoreactive silk sericin coupled with a conducting polymer. These functional electrodes are printed on flexible fibroin substrates that are controllably thick and can be free-standing, or conform to soft surfaces. Detection was accomplished via the antibody to VEGF which was immobilized within the conducting matrix. The results indicated that the developed flexible biosensor was highly sensitive and selective to the target protein, even in challenging biofluids such as human serum. The biosensors themselves are biocompatible and degradable. Through this work, the developed flexible biosensor based on a simple and label-free strategy can find practical applications in the monitoring of wound healing or early disease diagnosis.