Detection of Listeria monocytogenes in foods with a textile organic electrochemical transistor biosensor.

Foods contaminated by pathogens are responsible for foodborne diseases which have socioeconomic impacts. Many approaches have been extensively investigated to obtain specific and sensitive methods to detect pathogens in food, but they are often not easy to perform and require trained personnel. This work aims to propose a textile organic electrochemical transistor-based (OECT) biosensor to detect L. monocytogenes in food samples. The analyses were performed with culture-based methods, Listeria Precis™ method, PCR, and our textile OECT biosensor which used poly(3,4-ethylenedioxythiophene) (PEDOT):polystyrene sulfonate (PSS) (PEDOT:PSS) for doping the organic channel. Atomic force microscopy (AFM) was used to obtain topographic maps of the gold gate. The electrochemical activity on gate electrodes was measured and related to the concentration of DNA extracted from samples and hybridized to the specific capture probe immobilized onto the gold surface of the gate. This assay reached a limit of detection of 1.05 ng/µL, corresponding to 0.56 pM of L. monocytogenes ATCC 7644, and allowed the specific and rapid detection of L. monocytogenes in the analyzed samples. KEYPOINTS: • Textile organic electrochemical transistors functionalized with a specific DNA probe • AFM topographic and surface potential maps of a functionalized gold gate surface • Comparison between the Listeria monocytogenes Precis™ method and an OECT biosensor.

Long-Term Stability in Electronic Properties of Textile Organic Electrochemical Transistors for Integrated Applications.

Organic electrochemical transistors (OECTs) have demonstrated themselves to be an efficient interface between living environments and electronic devices in bioelectronic applications. The peculiar properties of conductive polymers allow new performances that overcome the limits of conventional inorganic biosensors, exploiting the high biocompatibility coupled to the ionic interaction. Moreover, the combination with biocompatible and flexible substrates, such as textile fibers, improves the interaction with living cells and allows specific new applications in the biological environment, including real-time analysis of plants' sap or human sweat monitoring. In these applications, a crucial issue is the lifetime of the sensor device. The durability, long-term stability, and sensitivity of OECTs were studied for two different textile functionalized fiber preparation processes: (i) adding ethylene glycol to the polymer solution, and (ii) using sulfuric acid as a post-treatment. Performance degradation was studied by analyzing the main electronic parameters of a significant number of sensors for a period of 30 days. RGB optical analysis were performed before and after the treatment of the devices. This study shows that device degradation occurs at voltages higher than 0.5 V. The sensors obtained with the sulfuric acid approach exhibit the most stable performances over time.

Methylglyoxal Adducts Levels in Blood Measured on Dried Spot by Portable Near-Infrared Spectroscopy.

The altered glucose metabolism characterising cancer cells determines an increased amount of methylglyoxal in their secretome. Previous studies have demonstrated that the methylglyoxal, in turn, modifies the protonation state (PS) of soluble proteins contained in the secretomes of cultivated circulating tumour cells (CTCs). In this study, we describe a method to assess the content of methylglyoxal adducts (MAs) in the secretome by near-infrared (NIR) portable handheld spectroscopy and the extreme learning machine (ELM) algorithm. By measuring the vibration absorption functional groups containing hydrogen, such as C-H, O-H and N-H, NIR generates specific spectra. These spectra reflect alterations of the energy frequency of a sample bringing information about its MAs concentration levels. The algorithm deciphers the information encoded in the spectra and yields a quantitative estimate of the concentration of MAs in the sample. This procedure was used for the comparative analysis of different biological fluids extracted from patients suspected of having cancer (secretome, plasma, serum, interstitial fluid and whole blood) measured directly on the solute left on a surface upon a sample-drop cast and evaporation, without any sample pretreatment. Qualitative and quantitative regression models were built and tested to characterise the different levels of MAs by ELM. The final model we selected was able to automatically segregate tumour from non-tumour patients. The method is simple, rapid and repeatable; moreover, it can be integrated in portable electronic devices for point-of-care and remote testing of patients.

Tailoring Chemometric Models on Blood-Derived Cultures Secretome to Assess Personalized Cancer Risk Score.

The molecular protonation profiles obtained by means of an organic electrochemical transistor, which is used for analysis of molecular products released by blood-derived cultures, contain a large amount of information The transistor is based on the conductive polymer PEDOT:PSS comprising super hydrophobic SU8 pillars positioned on the substrate to form a non-periodic square lattice to measure the state of protonation on secretomes derived from liquid biopsies. In the extracellular space of cultured cells, the number of glycation products increase, driven both by a glycolysis metabolism and by a compromised function of the glutathione redox system. Glycation products are a consequence of the interaction of the reactive aldehydes and side glycolytic products with other molecules. As a result, the amount of the glycation products reflects the anti-oxidative cellular reserves, counteracting the reactive aldehyde production of which both the secretome protonation profile and cancer risk are related. The protonation profiles can be profitably exploited through the use of mathematical techniques and multivariate statistics. This study provides a novel chemometric approach for molecular analysis of protonation and discusses the possibility of constructing a predictive cancer risk model based on the exploration of data collected by conventional analysis techniques and novel nanotechnological devices.

Emerging Designs of Electronic Devices in Biomedicine.

A long-standing goal of nanoelectronics is the development of integrated systems to be used in medicine as sensor, therapeutic, or theranostic devices. In this review, we examine the phenomena of transport and the interaction between electro-active charges and the material at the nanoscale. We then demonstrate how these mechanisms can be exploited to design and fabricate devices for applications in biomedicine and bioengineering. Specifically, we present and discuss electrochemical devices based on the interaction between ions and conductive polymers, such as organic electrochemical transistors (OFETs), electrolyte gated field-effect transistors (FETs), fin field-effect transistor (FinFETs), tunnelling field-effect transistors (TFETs), electrochemical lab-on-chips (LOCs). For these systems, we comment on their use in medicine.

Development of an In Vivo Sensor to Monitor the Effects of Vapour Pressure Deficit (VPD) Changes to Improve Water Productivity in Agriculture.

Environment, biodiversity and ecosystem services are essential to ensure food security and nutrition. Managing natural resources and mainstreaming biodiversity across agriculture sectors are keys towards a sustainable agriculture focused on resource efficiency. Vapour Pressure Deficit (VPD) is considered the main driving force of water movements in the plant vascular system, however the tools available to monitor this parameter are usually based on environmental monitoring. The driving motif of this paper is the development of an in-vivo sensor to monitor the effects of VPD changes in the plant. We have used an in vivo sensor, termed "bioristor", to continuously monitor the changes occurring in the sap ion's status when plants experience different VPD conditions and we observed a specific R (sensor response) trend in response to VPD. The possibility to directly monitor the physiological changes occurring in the plant in different VPD conditions, can be used to increase efficiency of the water management in controlled conditions thus achieving a more sustainable use of natural resources.

In Vivo Phenotyping for the Early Detection of Drought Stress in Tomato.

Drought stress imposes a major constraint over a crop yield and can be expected to grow in importance if the climate change predicted comes about. Improved methods are needed to facilitate crop management via the prompt detection of the onset of stress. Here, we report the use of an in vivo OECT (organic electrochemical transistor) sensor, termed as bioristor, in the context of the drought response of the tomato plant. The device was integrated within the plant's stem, thereby allowing for the continuous monitoring of the plant's physiological status throughout its life cycle. Bioristor was able to detect changes of ion concentration in the sap upon drought, in particular, those dissolved and transported through the transpiration stream, thus efficiently detecting the occurrence of drought stress immediately after the priming of the defence responses. The bioristor's acquired data were coupled with those obtained in a high-throughput phenotyping platform revealing the extreme complementarity of these methods to investigate the mechanisms triggered by the plant during the drought stress event.

An in vivo biosensing, biomimetic electrochemical transistor with applications in plant science and precision farming.

The in vivo monitoring of key plant physiology parameters will be a key enabler of precision farming. Here, a biomimetic textile-based biosensor, which can be inserted directly into plant tissue is presented: the device is able to monitor, in vivo and in real time, variations in the solute content of the plant sap. The biosensor has no detectable effect on the plant's morphology even after six weeks of continuous operation. The continuous monitoring of the sap electrolyte concentration in a growing tomato plant revealed a circadian pattern of variation. The biosensor has the potential to detect the signs of abiotic stress, and therefore might be exploited as a powerful tool to study plant physiology and to increase tomato growth sustainability. Also, it can continuously communicate the plant health status, thus potentially driving the whole farm management in the frame of smart agriculture.

Geometrical Patterning of Super-Hydrophobic Biosensing Transistors Enables Space and Time Resolved Analysis of Biological Mixtures.

PEDOT: PSS is a conductive polymer that can be integrated into last generation Organic Electrochemical Transistor (OECT) devices for biological inspection, identification and analysis. While a variety of reports in literature demonstrated the chemical and biological sensitivity of these devices, still their ability in resolving complex mixtures remains controversial. Similar OECT devices display good time dynamics behavior but lack spatial resolution. In this work, we integrated PEDOT: PSS with patterns of super-hydrophobic pillars in which a finite number of those pillars is independently controlled for site-selective measurement of a solution. We obtained a multifunctional, hierarchical OECT device that bridges the micro- to the nano-scales for specific, combined time and space resolved analysis of the sample. Due to super-hydrophobic surface properties, the biological species in the drop are driven by convection, diffusion, and the externally applied electric field: the balance/unbalance between these forces will cause the molecules to be transported differently within its volume depending on particle size thus realizing a size-selective separation. Within this framework, the separation and identification of two different molecules, namely Cetyl Trimethyl Ammonium Bromid (CTAB) and adrenaline, in a biological mixture have been demonstrated, showing that geometrical control at the micro-nano scale impart unprecedented selectivity to the devices.

Titanyl phthalocyanine ambipolar thin film transistors making use of carbon nanotube electrodes.

The capability of efficiently injecting charge carriers into organic films and finely tuning their morphology and structure is crucial to improve the performance of organic thin film transistors (OTFTs). In this work, we investigate OTFTs employing carbon nanotubes (CNTs) as the source-drain electrodes and, as the organic semiconductor, thin films of titanyl phthalocyanine (TiOPc) grown by supersonic molecular beam deposition (SuMBD). While CNT electrodes have shown an unprecedented ability to improve charge injection in OTFTs, SuMBD is an effective technique to tune film morphology and structure. Varying the substrate temperature during deposition, we were able to grow both amorphous (low substrate temperature) and polycrystalline (high substrate temperature) films of TiOPc. Regardless of the film morphology and structure, CNT electrodes led to superior charge injection and transport performance with respect to benchmark Au electrodes. Vacuum annealing of polycrystalline TiOPc films with CNT electrodes yielded ambipolar OTFTs.

Diffusion driven selectivity in organic electrochemical transistors.

Organic Electrochemical transistors (OECTs) present unique features for their strategic combination with biomedical interfaces, simple and low voltage operation regime and sensing ability in aqueous environment, but they still lack selectivity, so that a significant effort in research is devoted to overcome this limitation. Here, we focus on the diffusion properties of molecular species in the electrolyte, which opportunely analyzed, modeled and compared to experimental data, serve as a simple and direct key factor in the recognition of species during OECT sensing. Specifically, we model the transient behavior of an OECT considering the effect of diffusion of the target species in the electrolyte. In doing so, we develop a general method that can be used to differentiate and distinguish different molecules from a complex mixture, on the basis of their diffusivity and thus mass. More importantly, the model can be realistically used to determine the physical characteristics of the transported species in a solution from a simple fitting procedure. On the basis of the obtained results, we discuss the contribution that our study could bring to OECT architecture to realize a new generation of devices with improved sensitivity, selectivity and reliability.

Microtexturing of the conductive PEDOT:PSS polymer for superhydrophobic organic electrochemical transistors.

Superhydrophobic surfaces are bioinspired, nanotechnology artifacts, which feature a reduced friction coefficient, whereby they can be used for a number of very practical applications including, on the medical side, the manipulation of biological solutions. In this work, we integrated superhydrophobic patterns with the conducting polymer PEDOT:PSS, one of the most used polymers in organic electronics because highly sensitive to ionized species in solution. In doing so, we combined geometry and materials science to obtain an advanced device where, on account of the superhydrophobicity of the system, the solutions of interest can be manipulated and, on account of the conductive PEDOT:PSS polymer, the charged molecules dispersed inside can be quantitatively measured. This original substrate preparation allowed to perform electrochemical measurements on ionized species in solution with decreasing concentration down to 10(-7) molar. Moreover, it was demonstrated the ability of the device of realizing specific, combined time and space resolved analysis of the sample. Collectively, these results demonstrate how a tight, interweaving integration of different disciplines can provide realistic tools for the detection of pathologies. The scheme here introduced offers breakthrough capabilities that are expected to radically improve both the pace and the productivity of biomedical research, creating an access revolution.

Human stress monitoring through an organic cotton-fiber biosensor.

Selective detection of bioanalytes in physiological fluids, such as blood, sweat or saliva, by means of low-cost and non-invasive devices, is of crucial importance to improve diagnosis and prevention in healthcare. To be really useful in everyday life a sensing system needs to be handy, non-invasive, easy to read and possibly wearable. Only a sensor that satisfies these requirements could be eligible for applications in healthcare and physiological condition monitoring. Herein an organic electrochemical transistor has been investigated as a simple, low-cost and e-textile biosensor, fully integrated on a single cotton yarn. The biosensor has been used for real-time detection of adrenaline, selectively compared to the saline content in human physiological fluids. The sensing mechanism is based on the oxidation of adrenaline at the Pt-gate electrode surface, with the formation of adrenaline-quinone and adrenochrome. Two independent organic electrochemical transistors, characterized by different gate-electrode materials, detect saline and adrenaline concentrations, respectively, in real human sweat. Measurements performed in real-time mode show the complete independence of adrenaline detection from NaCl and, hence, guarantee the simultaneous monitoring of both concentrations. The oxidation of adrenaline has been studied by means of absorption spectroscopy in air, with either silver or platinum working electrodes. Our results confirm that the oxidation reaction driven by the Pt-electrode leads to the formation of adrenochrome, while with the Ag-electrode the oxidation is similar to the spontaneous one occurring in air. The cotton-based biosensor shows the possibility of monitoring human performances (hydration and stress) in situ and using a non-invasive approach, opening new unexplored opportunities in healthcare, fitness and work safety.

Low temperature sensing properties of a nano hybrid material based on ZnO nanotetrapods and titanyl phthalocyanine.

ZnO nanotetrapods have recently been exploited for the realization of high-sensitivity gas sensors, but they are affected by the typical drawbacks of metal-oxides, i.e., poor selectivity and a relatively high working temperature. On the other hand, it has been also demonstrated that the combined use of nanostructured metal oxides and organic molecules can improve the gas sensing performance sensitivity or selectivity, even at lower temperatures. A gas sensor device, based on films of interconnected ZnO nanotetrapods properly functionalized by titanyl phthalocyanine (TiOPc), has been realized in order to combine the high surface to volume ratio and structural stability of the crystalline ZnO nanostructures with the enhanced sensitivity of the semiconducting TiOPc molecule, especially at low temperature. The electronic properties of the resulting nanohybrid material are different from those of each single component. The response of the hybrid nanostructure towards different gases has been compared with that of ZnO nanotetrapod without functionalization in order to highlight the peculiar properties of the hybrid interaction(s). The dynamic response in time has been studied for different gases and temperatures; in particular, an increase in the response to NO2 has been observed, even at room temperature. The formation of localized p-n heterojunctions and the possibility of exchanging charge carriers at the hybrid interface is shown to be crucial for the sensing mechanism.

Liposome sensing and monitoring by organic electrochemical transistors integrated in microfluidics.

BACKGROUND: Organic electrochemical transistors (OECTs), which are becoming more and more promising devices for applications in bioelectronics and nanomedicine, are proposed here as ideally suitable for sensing and real time monitoring of liposome-based structures. This is quite relevant since, currently, the techniques used to investigate liposomal structures, their stability in different environments as well as drug loading and delivery mechanisms, operate basically off-line and/or with pre-prepared sampling. METHODS: OECTs, based on the PEDOT:PSS conductive polymer, have been employed as sensors of liposome-based nanoparticles in electrolyte solutions to assess sensitivity and monitoring capabilities based on ion-to-electron amplified transduction. RESULTS: We demonstrate that OECTs are very efficient, reliable and sensitive devices for detecting liposome-based nanoparticles on a wide dynamic range down to 10(-5)mg/ml (with a lowest detection limit, assessed in real-time monitoring, of 10(-7)mg/ml), thus matching the needs of typical drug loading/drug delivery conditions. They are hence particularly well suited for real-time monitoring of liposomes in solution. Furthermore, OECTs are shown to sense and discriminate successive injection of different liposomes, so that they could be good candidates in quality-control assays or in the pharmaceutical industry. GENERAL SIGNIFICANCE: Drug loading and delivery by liposome-based structures is a fast growing and very promising field that will strongly benefit from real-time, highly sensitive and low cost monitoring of their dynamics in different pharma and biomedical environments, with a particular reference to the pharmaceutical and production processes, where a major issue is monitoring and measuring the formation and concentration of liposomes and the relative drug load. The demonstrated ability to sense and monitor complex bio-structures, such as liposomes, paves the way for very promising developments in biosensing and nanomedicine. This article is part of a Special Issue entitled Organic Bioelectronics-Novel Applications in Biomedicine.

Irreversible evolution of eumelanin redox states detected by an organic electrochemical transistor: en route to bioelectronics and biosensing.

Organic electrochemical transistors (OECTs) are currently emerging as powerful tools for biosensing, bioelectronics and nanomedical applications owing to their ability to operate under liquid phase conditions optimally integrating electronic and biological systems. Herein we disclose the unique potential of OECTs for detecting and investigating the electrical properties of insoluble eumelanin biopolymers. Gate current measurements on fine aqueous suspensions of a synthetic eumelanin sample from 5,6-dihydroxyindole (DHI) revealed a well detectable hysteretic response similar to that of the pure monomer in solution, with the formal concentration of the polymer as low as 10-6 M. Induction of the gate current would reflect electron transfer from solid eumelanin to the Pt-electrode sustained by redox active catechol/quinone components of the polymer. A gradual decrease in gate current and areas subtended by hysteretic loops were observed over 5 cycles both in the eumelanin- and DHI-based devices, suggesting evolution of the polymer from a far-from-the-equilibrium redox state toward a more stable electronic arrangement promoted by redox exchange with the gate electrode. OECTs are thus proposed as valuable tools for the efficient heterogeneous-phase sensing of eumelanins and to gauge their peculiar electrical and redox behaviour.

Superhydrophobic surfaces as smart platforms for the analysis of diluted biological solutions.

The aim of this paper is to expound on the rational design, fabrication and development of superhydrophobic surfaces (SHSs) for the manipulation and analysis of diluted biological solutions. SHSs typically feature a periodic array or pattern of micropillars; here, those pillars were modified to incorporate on the head, at the smallest scales, silver nanoparticles aggregates. These metal nanoclusters guarantee superior optical properties and especially SERS (surface enhanced Raman scattering) effects, whereby a molecule, adsorbed on the surface, would reveal an increased spectroscopy signal. On account of their two scale-hybrid nature, these systems are capable of multiple functions which are (i) to concentrate a solution, (ii) to vehicle the analytes of interest to the active areas of the substrate and, therefore, (iii) to measure the analytes with exceptional sensitivity and very low detection limits. Forasmuch, combining different technologies, these devices would augment the performance of conventional SERS substrates and would offer the possibility of revealing a single molecule. In this work, similar SHSs were used to detect Rhodamine molecules in the fairly low atto molar range. The major application of this novel family of devices would be the early detection of tumors or other important pathologies, with incredible advances in medicine.

Polymorphism and phase control in titanyl phthalocyanine thin films grown by supersonic molecular beam deposition.

Polymorphism in the growth of titanyl phthalocyanine films on dielectric substrates has been systematically studied by UV absorption and micro-Raman analyses, correlating structure and optical properties. We explored different growth regimes as a function of substrate temperature and growth rate using hyperthermal seeded supersonic beams. We identify and discuss specific signatures in micro-Raman spectra specifically correlated to the different phases and demonstrate the unprecedented ability of growing crystalline films and controlling the relative abundance of the different phases (amorphous, phase I, and phase II) by the beam parameters. We envisage the very promising perspective of controlling polymorphism at low temperatures via supersonic beam growth, paving the way for better performing devices.