Craft-and-Stick Xurographic Manufacturing of Integrated Microfluidic Electrochemical Sensing Platform.

An innovative modular approach for facile design and construction of flexible microfluidic biosensor platforms based on a dry manufacturing "craft-and-stick" approach is developed. The design and fabrication of the flexible graphene paper electrode (GPE) unit and polyethylene tetraphthalate sheet (PET)6/adhesive fluidic unit are completed by an economic and generic xurographic craft approach. The GPE widths and the microfluidic channels can be constructed down to 300 µm and 200 µm, respectively. Both units were assembled by simple double-sided adhesive tapes into a microfluidic integrated GPE (MF-iGPE) that are flexible, thin (<0.5 mm), and lightweight (0.4 g). We further functionalized the iGPE with Prussian blue and glucose oxidase for the fabrication of MF-iGPE glucose biosensors. With a closed-channel PET fluidic pattern, the MF-iGPE glucose biosensors were packaged and sealed to protect the integrated device from moisture for storage and could easily open with scissors for sample loading. Our glucose biosensors showed 2 linear dynamic regions of 0.05-1.0 and 1.0-5.5 mmol L-1 glucose. The MF-iGPE showed good reproducibility for glucose detection (RSD < 6.1%, n = 6) and required only 10 µL of the analyte. This modular craft-and-stick manufacturing approach could potentially further develop along the concept of paper-crafted model assembly kits suitable for low-resource laboratories or classroom settings.

A portable smartphone-based imaging surface plasmon resonance biosensor for allergen detection in plant-based milks.

Food allergies are hypersensitivity immune responses triggered by (traces of) allergenic compounds in foods and drinks. The recent trend towards plant-based and lactose-free diets has driven an increased consumption of plant-based milks (PBMs) with the risk of cross-contamination of various allergenic plant-based proteins during the food manufacturing process. Conventional allergen screening is usually performed in the laboratory, but portable biosensors for on-site screening of food allergens at the production site could improve quality control and food safety. Here, we developed a portable smartphone imaging surface plasmon resonance (iSPR) biosensor composed of a 3D-printed microfluidic SPR chip for the detection of total hazelnut protein (THP) in commercial PBMs and compared its instrumentation and analytical performance with a conventional benchtop SPR. The smartphone iSPR shows similar characteristic sensorgrams compared with the benchtop SPR and enables the detection of trace levels of THP in spiked PBMs with the lowest tested concentration of 0.625 µg/mL THP. The smartphone iSPR achieved LoDs of 0.53, 0.16, 0.14, 0.06, and 0.04 µg/mL THP in 10x-diluted soy, oat, rice, coconut, and almond PBMs, respectively, with good correlation with the conventional benchtop SPR system (R2 0.950-0.991). The portability and miniaturized characteristics of the smartphone iSPR biosensor platform make it promising for the future on-site detection of food allergens by food producers.

A wireless smartphone-based "tap-and-detect" formaldehyde sensor with disposable nano-palladium grafted laser-induced graphene (nanoPd@LIG) electrodes.

We developed a fully integrated smart sensing device for on-site testing of food to detect trace formaldehyde (FA). A nano-palladium grafted laser-induced graphene (nanoPd@LIG) composite was synthesized by one-step laser irradiation of a Pd2+-chitosan-polyimide precursor. The composite was synthesized in the form of a three-electrode sensor on a polymer substrate. The electrochemical properties and morphology of the fabricated composite were characterized and the electrochemical kinetics of FA oxidation at the nanoPd@LIG electrode were investigated. The nanoPd@LIG electrode was combined with a smart electrochemical sensing (SES) device to determine FA electrochemically. The proposed SES device uses near field communication (NFC) to receive power and transfer data between a smartphone interface and a battery-free sensor. The proposed FA sensor exhibited a linear detection range from 0.01 to 4.0 mM, a limit of detection of 6.4 µM, good reproducibility (RSDs between 2.0 and 10.1%) and good anti-interference properties for FA detection. The proposed system was used to detect FA in real food samples and the results correlated well with the results from a commercial potentiostat and a spectrophotometric analysis.

An Integrated ddPCR Lab-on-a-Disc Device for Rapid Screening of Infectious Diseases.

Digital droplet PCR (ddPCR) is a powerful amplification technique for absolute quantification of viral nucleic acids. Although commercial ddPCR devices are effective in the lab bench tests, they cannot meet current urgent requirements for on-site and rapid screening for patients. Here, we have developed a portable and fully integrated lab-on-a-disc (LOAD) device for quantitively screening infectious disease agents. Our designed LOAD device has integrated (i) microfluidics chips, (ii) a transparent circulating oil-based heat exchanger, and (iii) an on-disc transmitted-light fluorescent imaging system into one compact and portable box. Thus, droplet generation, PCR thermocycling, and analysis can be achieved in a single LOAD device. This feature is a significant attribute for the current clinical application of disease screening. For this custom-built ddPCR setup, we have first demonstrated the loading and ddPCR amplification ability by using influenza A virus-specific DNA fragments with different concentrations (diluted from the original concentration to 107 times), followed by analyzing the droplets with an external fluorescence microscope as a standard calibration test. The measured DNA concentration is linearly related to the gradient-dilution factor, which validated the precise quantification for the samples. In addition to the calibration tests using DNA fragments, we also employed this ddPCR-LOAD device for clinical samples with different viruses. Infectious samples containing five different viruses, including influenza A virus (IAV), respiratory syncytial virus (RSV), varicella zoster virus (VZV), Zika virus (ZIKV), and adenovirus (ADV), were injected into the device, followed by analyzing the droplets with an external fluorescence microscope with the lowest detected concentration of 20.24 copies/µL. Finally, we demonstrated the proof-of-concept detection of clinical samples of IAV using the on-disc fluorescence imaging system in our fully integrated device, which proves the capability of this device in clinical sample detection. We anticipate that this integrated ddPCR-LOAD device will become a flexible tool for on-site disease detection.

A green route for lignin-derived graphene electrodes: A disposable platform for electrochemical biosensors.

The tremendous growth of disposable electrode-based portable devices for point-of-care testing requires mass production of disposable electrodes in a low-cost and sustainable manner. Here, we demonstrate a green route for the conversion of biomass lignin, patterning, and reduction of the lignin-derived graphene electrodes by sequential laser lithography, water lift-off and sodium borohydride (NaBH4) treatment, and their use for electrochemical lactate biosensors. Energy-saving and localized laser lithography converted the aromatic ring-rich lignin into porous laser-induced graphene (LIG). The conductivity and attachment of the LIG to the substrate were optimized in a factorial experiment with laser power and scan speed as variables. Characterization results revealed the conversion of partial heteroatoms (e.g., Na, S, O) into granular inorganic compounds on the LIG surface under laser treatment. Water was used as an eco-friendly solvent for the patterning of the LIG (P-LIG) by a lift-off process, where the inorganic residues and un-reacted lignin were dissolved, exposing the macro-/micro-pores in the P-LIG. NaBH4 induced a reduction of the P-LIG (P-rLIG) resulting in improved electrochemical kinetics with lower charge transfer resistance (27.3 Ω) compared to the LIG (248.1 Ω) and the P-LIG (61.4 Ω). The porous P-rLIG served as a 3D electrode for the deposition of Prussian blue and lactate oxidase for disposable electrochemical lactate biosensors, delivering a good analytical performance towards lactate detection with a linear range up to 16 mM and a high sensitivity (1.21 µA mM-1). These lignin-derived disposable electrodes, utilizing renewable resources together with low-energy consumption fabrication and patterning, may contribute to the sustainable manufacturing of biosensors for point-of-care and point-of-use applications.

Low-cost and rapid prototyping of integrated electrochemical microfluidic platforms using consumer-grade off-the-shelf tools and materials.

We present a low-cost, accessible, and rapid fabrication process for electrochemical microfluidic sensors. This work leverages the accessibility of consumer-grade electronic craft cutters as the primary tool for patterning of sensor electrodes and microfluidic circuits, while commodity materials such as gold leaf, silver ink pen, double-sided tape, plastic transparency films, and fabric adhesives are used as its base structural materials. The device consists of three layers, the silver reference electrode layer at the top, the PET fluidic circuits in the middle and the gold sensing electrodes at the bottom. Separation of the silver reference electrode from the gold sensing electrodes reduces the possibility of cross-contamination during surface modification. A novel approach in mesoscale patterning of gold leaf electrodes can produce generic designs with dimensions as small as 250 µm. Silver electrodes with dimensions as small as 385 µm were drawn using a plotter and a silver ink pen, and fluid microchannels as small as 300 µm were fabricated using a sandwich of iron-on adhesives and PET. Device layers are then fused together using an office laminator. The integrated microfluidic electrochemical platform has electrode kinetics/performance of ΔEp = 91.3 mV, Ipa/Ipc = 0.905, characterized by cyclic voltammetry using a standard ferrocyanide redox probe, and this was compared against a commercial screen-printed gold electrode (ΔEp = 68.9 mV, Ipa/Ipc = 0.984). To validate the performance of the integrated microfluidic electrochemical platform, a catalytic hydrogen peroxide sensor and enzyme-coupled glucose biosensors were developed as demonstrators. Hydrogen peroxide quantitation achieves a limit of detection of 0.713 mM and sensitivity of 78.37 µA mM-1 cm-2, while glucose has a limit of detection of 0.111 mM and sensitivity of 12.68 µA mM-1 cm-2. This rapid process allows an iterative design-build-test cycle in under 2 hours. The upfront cost to set up the system is less than USD 520, with each device costing less than USD 0.12, making this manufacturing process suitable for low-resource laboratories or classroom settings.

Print-and-stick unibody microfluidics coupled surface plasmon resonance (SPR) chip for smartphone imaging SPR (Smart-iSRP).

The design of a smartphone imaging surface plasmon resonance (Smart-iSPR) system integrated with an affordable 3D-printed microfluidic SPR chip fabricated via a facile manufacturing approach could pave the way towards the development of miniaturized and integrated smartphone iSPR biosensors for emerging point-of-use applications. Conventional smartphone-based SPR systems using soft photolithography for the fabrication of microfluidics SPR chips are costly, labour-intensive and required a specially-equipped light-controlled environment, that is inadequate and mismatched with the consumer-based smartphone detection platform. Herein, we report the design, fabrication and testing of an innovative print-and-stick unibody microfluidics coupled SPR chip for smartphone iSPR. The 3D-printed microfluidics (∼€0.006) is assembled via an aptly-sized adhesive tape with the gold SPR sensing surface. Such a simple integrated microfluidic SPR chip with the print-and-stick configuration has a high resistance to fluid leakages at the channel-to-sensor interface with pressure up to 66.9 Pa and the tubing-to-inset interfaces with pressure up to 86.9 Pa. The smartphone iSPR platform weighs 138 g and with a dimension of around 70 × 60 × 40 mm3, and its performance was characterized using a standard Biacore® ß2-microglobulin calibration kit. The sensorgrams obtained by the smartphone iSPR show all the typical characteristics for surface functionalization, association and dissociation events. The smartphone iSPR responds linearly to ß2-microglobulin within the range of 10-200 nM (R2 = 0.986) with a limit-of-detection (LOD) of 1.5 nM. Given the miniaturized feature and simple camera-based imaging smartphone iSPR, the analytical performance is satisfactory when compared with the analytical dynamic range of 2-32 nM described in the Biacore® protocol.

Evaluation on the Intrinsic Physicoelectrochemical Attributes and Engineering of Micro-, Nano-, and 2D-Structured Allotropic Carbon-Based Papers for Flexible Electronics.

Flexible electronics have gained more attention for emerging electronic devices such as sensors, biosensors, and batteries with advantageous properties including being thin, lightweight, flexible, and low-cost. The development of various forms of allotropic carbon papers provided a new dry-manufacturing route for the fabrication of flexible and wearable electronics, while the electrochemical performance and the bending stability are largely influenced by the bulk morphology and the micro-/nanostructured domains of the carbon papers. Here, we evaluate systematically the intrinsic physicoelectrochemical properties of allotropic carbon-based conducting papers as flexible electrodes including carbon-nanotubes-paper (CNTs-paper), graphene-paper (GR-paper), and carbon-fiber-paper (CF-paper), followed by functionalization of the allotropic carbon papers for the fabrication of flexible electrodes. The morphology, chemical structure, and defects originating from the allotropic nanostructured carbon materials were characterized by scanning electron microscopy (SEM) and Raman spectroscopy, followed by evaluating the electrochemical performance of the corresponding flexible electrodes by cyclic voltammetry and electrochemical impedance spectroscopy. The electron-transfer rate constants of the CNTs-paper and GR-paper electrodes were ∼14 times higher compared with the CF-paper electrode. The CNTs-paper and GR-paper electrodes composed of nanostructured carbon showed significantly higher bending stabilities of 5.61 and 4.96 times compared with the CF-paper. The carbon-paper flexible electrodes were further functionalized with an inorganic catalyst, Prussian blue (PB), forming the PB-carbon-paper catalytic electrode and an organic conducting polymer, poly(3,4-ethylenedioxythiophene) (PEDOT), forming the PEDOT-carbon-paper capacitive electrode. The intrinsic attribute of different allotropic carbon electrodes affects the deposition of PB and PEDOT, leading to different electrocatalytic and capacitive performances. These findings are insightful for the future development and fabrication of advanced flexible electronics with allotropic carbon papers.

RF Remote Blood Glucose Sensor and a Microfluidic Vascular Phantom for Sensor Validation.

Diabetes has become a major health problem in society. Invasive glucometers, although precise, only provide discrete measurements at specific times and are unsuitable for long-term monitoring due to the injuries caused on skin and the prohibitive cost of disposables. Remote, continuous, self-monitoring of blood sugar levels allows for active and better management of diabetics. In this work, we present a radio frequency (RF) sensor based on a stepped impedance resonator for remote blood glucose monitoring. When placed on top of a human hand, this RF interdigital sensor allows detection of variation in blood sugar levels by monitoring the changes in the dielectric constant of the material underneath. The designed stepped impedance resonator operates at 3.528 GHz with a Q factor of 1455. A microfluidic device structure that imitates the blood veins in the human hand was fabricated in PDMS to validate that the sensor can measure changes in glucose concentrations. To test the RF sensor, glucose solutions with concentrations ranging from 0 to 240 mg/dL were injected into the fluidic channels and placed underneath the RF sensor. The shifts in the resonance frequencies of the RF sensor were measured using a network analyzer via its S11 parameters. Based on the change in resonance frequencies, the sensitivity of the biosensor was found to be 264.2 kHz/mg·dL-1 and its LOD was calculated to be 29.89 mg/dL.

Conducting Polymer-Reinforced Laser-Irradiated Graphene as a Heterostructured 3D Transducer for Flexible Skin Patch Biosensors.

Flexible skin patch biosensors are promising for the noninvasive determination of physiological parameters in perspiration for fitness and health monitoring. However, various prerequisites need to be met for the development of such biosensors, including the creation of a flexible conductive platform, bending/contact stability, fast electrochemical kinetics, and immobilization of biomolecules. Here, we describe a conducting polymer-reinforced laser-irradiated graphene (LIG) network as a heterostructured three-dimensional (3D) transducer for flexible skin patch biosensors. LIG with a hierarchically interconnected graphene structure is geometrically patterned on polyimide via localized laser irradiation as a flexible conductive platform, which is then reinforced by poly(3,4-ethylenedioxythiophene) (PEDOT) as a conductive binder (PEDOT/LIG) with improved structural/contact stability and electrochemical kinetics. The interconnected pores of the reinforced PEDOT/LIG function as a 3D host matrix for high loading of "artificial" (Prussian blue, PB) and natural enzymes (lactate oxidase, LOx), forming a compact and heterostructured 3D transducer (LOx/PB-PEDOT/LIG) for lactate biosensing with excellent sensitivity (11.83 µA mM-1). We demonstrated the fabrication of flexible skin patch biosensors comprising a custom-built integrated three-electrode system achieve amperometric detection of lactate in artificial sweat over a wide physiological linear range of 0-18 mM. The advantage of this facile and versatile transducer is further illustrated by the development of a folded 3D wristband lactate biosensor and a dual channel biosensors for simultaneous monitoring of lactate and glucose. This innovative design concept of a heterostructured transducer for flexible biosensors combined with a versatile fabrication approach could potentially drive the development of new wearable and skin-mountable biosensors for monitoring various physiological parameters in biofluids for noninvasive fitness and health management.

Spatiotemporal extracellular matrix modeling for in situ cell niche studies.

Extracellular matrix (ECM) components govern a range of cell functions, such as migration, proliferation, maintenance of stemness, and differentiation. Cell niches that harbor stem-/progenitor cells, with matching ECM, have been shown in a range of organs, although their presence in the heart is still under debate. Determining niches depends on a range of in vitro and in vivo models and techniques, where animal models are powerful tools for studying cell-ECM dynamics; however, they are costly and time-consuming to use. In vitro models based on recombinant ECM proteins lack the complexity of the in vivo ECM. To address these issues, we present the spatiotemporal extracellular matrix model for studies of cell-ECM dynamics, such as cell niches. This model combines gentle decellularization and sectioning of cardiac tissue, allowing retention of a complex ECM, with recellularization and subsequent image processing using image stitching, segmentation, automatic binning, and generation of cluster maps. We have thereby developed an in situ representation of the cardiac ECM that is useful for assessment of repopulation dynamics and to study the effect of local ECM composition on phenotype preservation of reseeded mesenchymal progenitor cells. This model provides a platform for studies of organ-specific cell-ECM dynamics and identification of potential cell niches.

Precise and rapid solvent-assisted geometric protein self-patterning with submicron spatial resolution for scalable fabrication of microelectronic biosensors.

Precise and high-resolution coupling of functional proteins with micro-transducers is critical for the manufacture of miniaturized bioelectronic devices. Moreover, electrochemistry on microelectrodes has had a major impact on electrochemical analysis and sensor technologies, since the small size of microelectrode affects the radial diffusion flux of the analyte to deliver enhanced mass transport and electrode kinetics. However, a large technology gap has existed between the process technology associated with such microelectronics and the conventional bio-conjugation techniques that are generally used. Here, we report on a high-resolution and rapid geometric protein self-patterning (GPS) method using solvent-assisted protein-micelle adsorption printing to couple biomolecules onto microelectrodes with a minimum feature size of 5 µm and a printing time of about a minute. The GPS method is versatile for micropatterning various biomolecules including enzymes, antibodies and avidin-biotinylated proteins, delivering good geometric alignment and preserving biological functionality. We further demonstrated that enzyme-coupled microelectrodes for glucose detection exhibited good electrochemical performance which benefited from the GPS method to maximize effective signal transduction at the bio-interface. These microelectrode arrays maintained fast convergent analyte diffusion displaying typical steady-state I-V characteristics, fast response times, good linear sensitivity (0.103 nA mm-2 mM-1, R2 = 0.995) and an ultra-wide linear dynamic range (2-100 mM). Our findings provide a new technical solution for the precise and accurate coupling of biomolecules to a microelectronic array with important implications for the scaleup and manufacture of diagnostics, biofuel cells and bioelectronic devices that could not be realized economically by other existing techniques.

Processable and nanofibrous polyaniline:polystyrene-sulphonate (nano-PANI:PSS) for the fabrication of catalyst-free ammonium sensors and enzyme-coupled urea biosensors.

Tailoring conducting polymers (CPs) such as polyaniline (PANI) to deliver the appropriate morphology, electrochemical properties and processability is essential for the development of effective polymer-based electrochemical sensors and biosensors. Composite PANI electrodes for the detection of ammonium (NH4+) have been previously reported, but have been limited by their reliance on the electrocatalytic reaction between NH4+ and a metal/nano-catalyst. We report an advanced processable and nanofibrous polyaniline:polystyrene-sulphonate (nano-PANI:PSS) as a functional ink for the fabrication of catalyst-free NH4+ sensors and enzyme-coupled urea biosensors. The PSS provides both a soft-template for nanofibre formation and a poly-anionic charge compensator, enabling the detection of NH4+ based on an intrinsic doping/de-doping mechanism. The nanostructured morphology, chemical characteristics and electrochemical properties of the nano-PANI:PSS were characterised. We fabricated 3D-hierarchical sensor interfaces composed of inter-connected nano-PANI:PSS fibres (diameter of ~50.3 ± 4.8 nm) for the detection of NH4+ with a wide linear range of 0.1-11.5 mM (R2 = 0.996) and high sensitivity of 106 mA M-1 cm-2. We further demonstrated the coupling of the enzyme urease with the nano-PANI:PSS to create a urea biosensor with an innovative biocatalytic product-to-dopant relay mechanism for the detection of urea, with a linear range of 0.2-0.9 mM (R2 = 0.971) and high sensitivity of 41 mA M-1 cm-2. Moreover, the nano-PANI:PSS-based sensors show good selectivity for the detection of NH4+and urea in a urine model containing common interfering molecules. This processable and fibrous nano-PANI:PSS provides new advance on CP-based transducer materials in the emerging field of printed organic sensors and biosensors.

Bio-PEDOT: Modulating Carboxyl Moieties in Poly(3,4-ethylenedioxythiophene) for Enzyme-Coupled Bioelectronic Interfaces.

Modulation of chemical functional groups on conducting polymers (CPs) provides an effective way to tailor the physicochemical properties and electrochemical performance of CPs, as well as serves as a functional interface for stable integration of CPs with biomolecules for organic bioelectronics (OBEs). Herein, we introduced a facile approach to modulate the carboxylate functional groups on the PEDOT interface through a systematic evaluation on the effect of a series of carboxylate-containing molecules as counterion dopant integrated into the PEDOT backbone, including acetate as monocarboxylate (mono-COO-), malate as dicarboxylate (di-COO-), citrate as tricarboxylate (tri-COO-), and poly(acrylamide-co-acrylate) as polycarboxylate (poly-COO-) bearing different amounts of molecular carboxylate moieties to create tunable PEDOT:COO- interfaces with improved polymerization efficiency. We demonstrated the modulation of PEDOT:COO- interfaces with various granulated morphologies from 0.33 to 0.11 µm, tunable surface carboxylate densities from 0.56 to 3.6 µM cm-2, and with improved electrochemical kinetics and cycling stability. We further demonstrated the effective and stable coupling of an enzyme model lactate dehydrogenase (LDH) with the optimized PEDOT:poly-COO- interface via simple covalent chemistry to develop biofunctionalized PEDOT (Bio-PEDOT) as a lactate biosensor. The biosensing mechanism is driven by a sequential bioelectrochemical signal transduction between the bio-organic LDH and organic PEDOT toward the concept of all-polymer-based OBEs with a high sensitivity of 8.38 µA mM-1 cm-2 and good reproducibility. Moreover, we utilized the LDH-PEDOT biosensor for the detection of lactate in spiked serum samples with a high recovery value of 91-96% and relatively small RSD in the range of 2.1-3.1%. Our findings provide a new insight into the design and optimization of functional CPs, leading to the development of new OBEs for sensing, biosensing, bioengineering, and biofuel cell applications.

Tunable 3D nanofibrous and bio-functionalised PEDOT network explored as a conducting polymer-based biosensor.

Conducting polymers that possess good electrochemical properties, nanostructured morphology and functionality for bioconjugation are essential to realise the concept of all-polymer-based biosensors that do not depend on traditional nanocatalysts such as carbon materials, metal, metal oxides or dyes. In this research, we demonstrated a facile approach for the simultaneous preparation of a bi-functional PEDOT interface with a tunable 3D nanofibrous network and carboxylic acid groups (i.e. Nano-PEDOT-COOH) via controlled co-polymerisation of EDOT and EDOT-COOH monomers, using tetrabutylammonium perchlorate as a soft-template. By tuning the ratio between EDOT and EDOT-COOH monomer, the nanofibrous structure and carboxylic acid functionalisation of Nano-PEDOT-COOH were varied over a fibre diameter range of 15.6 ± 3.7 to 70.0 ± 9.5 nm and a carboxylic acid group density from 0.03 to 0.18 µmol cm-2. The nanofibres assembled into a three-dimensional network with a high specific surface area, which contributed to low charge transfer resistance and high transduction activity towards the co-enzyme NADH, delivering a wide linear range of 20-960 µM and a high sensitivity of 0.224 µA µM-1 cm-2 at the Nano-PEDOT-COOH50% interface. Furthermore, the carboxylic acid groups provide an anchoring site for the stable immobilisation of an NADH-dependent dehydrogenase (i.e. lactate dehydrogenase), via EDC/S-NHS chemistry, for the fabrication of a Bio-Nano-PEDOT-based biosensor for lactate detection which had a response time of less than 10 s over the range of 0.05-1.8 mM. Our developed bio-Nano-PEDOT interface shows future potential for coupling with multi-biorecognition molecules via carboxylic acid groups for the development of a range of advanced all-polymer biosensors.

Soft and flexible material-based affinity sensors.

Recent advances in biosensors and point-of-care (PoC) devices are poised to change and expand the delivery of diagnostics from conventional lateral-flow assays and test strips that dominate the market currently, to newly emerging wearable and implantable devices that can provide continuous monitoring. Soft and flexible materials are playing a key role in propelling these trends towards real-time and remote health monitoring. Affinity biosensors have the capability to provide for diagnosis and monitoring of cancerous, cardiovascular, infectious and genetic diseases by the detection of biomarkers using affinity interactions. This review tracks the evolution of affinity sensors from conventional lateral-flow test strips to wearable/implantable devices enabled by soft and flexible materials. Initially, we highlight conventional affinity sensors exploiting membrane and paper materials which have been so successfully applied in point-of-care tests, such as lateral-flow immunoassay strips and emerging microfluidic paper-based devices. We then turn our attention to the multifarious polymer designs that provide both the base materials for sensor designs, such as PDMS, and more advanced functionalised materials that are capable of both recognition and transduction, such as conducting and molecularly imprinted polymers. The subsequent content discusses wearable soft and flexible material-based affinity sensors, classified as flexible and skin-mountable, textile materials-based and contact lens-based affinity sensors. In the final sections, we explore the possibilities for implantable/injectable soft and flexible material-based affinity sensors, including hydrogels, microencapsulated sensors and optical fibers. This area is truly a work in progress and we trust that this review will help pull together the many technological streams that are contributing to the field.

Modulating Electrode Kinetics for Discrimination of Dopamine by a PEDOT:COOH Interface Doped with Negatively Charged Tricarboxylate.

The rapidly developing field of conducting polymers in organic electronics has many implications for bioelectronics. For biosensing applications, tailoring the functionalities of the conducting polymer's surface is an efficient approach to improve both sensitivity and selectivity. Here, we demonstrated a facile and economic approach for the fabrication of a high-density, negatively charged carboxylic-acid-group-functionalized PEDOT (PEDOT:COOH) using an inexpensive ternary carboxylic acid, citrate, as a dopant. The polymerization efficiency was significantly improved by the addition of LiClO4 as a supporting electrolyte yielding a dense PEDOT:COOH sensing interface. The resulting PEDOT:COOH interface had a high surface density of carboxylic acid groups of 0.129 µmol/cm2 as quantified by the toluidine blue O (TBO) staining technique. The dopamine response measured with the PEDOT:COOH sensing interface was characterized by cyclic voltammetry with a significantly reduced ΔEp of 90 mV and a 3-fold increase in the Ipa value compared with those of the nonfunctionalized PEDOT sensing interface. Moreover, the cyclic voltammetry and electrochemical impedance spectroscopy results demonstrated the increased electrode kinetics and highly selective discrimination of dopamine (DA) in the presence of the interferents ascorbic acid (AA) and uric acid (UA), which resulted from the introduction of negatively charged carboxylic acid groups. The negatively charged carboxylic acid groups could favor the transfer, preconcentration, and permeation of positively charged DA to deliver improved sensing performance while repelling the negatively charged AA and UA interferents. The PEDOT:COOH interface facilitated measurement of dopamine over the range of 1-85 µM, with a sensitivity of 0.228 µA µM-1, which is 4.1 times higher than that of a nonfunctionalized PEDOT electrode (0.055 µA µM-1). Our results demonstrate the feasibility of a simple and economic fabrication of a high-density PEDOT:COOH interface for chemical sensing, which also has the potential for coupling with other biorecognition molecules via carboxylic acid moieties for the development of a range of advanced PEDOT-based biosensors.

Integrated Printed Microfluidic Biosensors.

Integrated printed microfluidic biosensors are one of the most recent point-of-care (POC) sensor developments. Fast turnaround time for production and ease of customization, enabled by the integration of recognition elements and transducers, are key for on-site biosensing for both healthcare and industry and for speeding up translation to real-life applications. Here, we provide an overview of recent progress in printed microfluidics, from the 2D to the 4D level, accompanied by novel sensing element integration. We also explore the latest trends in integrated printed microfluidics for healthcare, especially POC diagnostics, and food safety applications.

Generic Neutravidin Biosensor for Simultaneous Multiplex Detection of MicroRNAs via Electrochemically Encoded Responsive Nanolabels.

Current electrochemical biosensors for multiple miRNAs require tedious immobilization of various nucleic acid probes. Here, we demonstrate an innovative approach using a generic neutravidin biosensor combined with electrochemically encoded responsive nanolabels for facile and simultaneous multiplexed detection of miRNA-21 and miRNA-141. The selectivity of the biosensor arises from the intrinsic properties of the electrochemically encoded responsive nanolabels, comprising biotinylated molecular beacons (biotin-MB) and metal nanoparticles (metal-NPs). The procedure is a simple one-pot assay, where the targeted miRNA causes the opening of biotin-MB followed by capturing of the biotin-MB-metal-NPs by the neutravidin biosensor and simultaneous detection of the captured metal-NPs by stripping square-wave voltammetry (SSWV). The multiplexed detection of miRNA-21 and miRNA-141 is achieved by differentiation of the electrochemical signature (i.e., the peak current) for the different metal-NP labels. The biosensor delivers simultaneous detection of miRNAs with a linear range of 0.5-1000 pM for miRNA-21 and a limit of detection of 0.3 pM (3σ/sensitivity,  n = 3), and a range of 50-1000 pM for miRNA-141, with a limit of detection of 10 pM. Furthermore, we demonstrate multiplexed detection of miRNA-21 and miRNA-141 in a spiked serum sample.

Integrating an ex-vivo skin biointerface with electrochemical DNA biosensor for direct measurement of the protective effect of UV blocking agents.

Skin cancer is the most frequent kind of cancer in white people in many parts of the world. UV-induced DNA damage and genetic mutation can subsequently lead to skin cancer. Therefore development of new biosensing strategies for detection of UV-induced DNA damage is of great importance. Here we demonstrate a novel combination of an ex-vivo skin biointerface and an electrochemical DNA sensor for the direct detection of UV induced DNA damage and investigation the protective effect of various UV blockers (Zinc-oxide (ZnO), titanium-dioxide (TiO2) nanoparticles (NPs) and sunscreens) against DNA damage. A diazonium modified screen-printed carbon electrode immobilized with a DNA sequence related to the p53 tumour suppressor gene, the most commonly affected gene in human UV-induced skin cancer, was applied as an electrochemical DNA sensor. Electrochemical impedance spectroscopy (EIS) was employed for the detection of DNA damage induced by UV-A radiation by following the changes in charge transfer resistance (Rct). The protective effects of UV blockers applied onto a pig skin surface (a suitable model representing human skin) were successfully detected by the DNA sensor. We observed that the naked skin has little UV protection showing an 18.2% decreases in ∆R/R values compared to the control, while applying both NPs and NP-formulated sunscreens could significantly reduce DNA damage, resulting in a decrease in ∆R/R values of 67.1% (ZnO NPs), 77.2% (TiO2 NPs), 77.1% (sunscreen 1) and 92.4% (sunscreen 2), respectively. Moreover, doping moisturising cream with NPs could provide a similar DNA protective effect. This new method is a biologically relevant alternative to animal testing and offers advantages such as fast, easy and inexpensive processing, in addition to its miniaturised dimension, and could be used for a range of applications in other sources of DNA damage and the protective effect of different UV blocking agents and other topical formulations.