Large-Area Interfaces for Single-Molecule Label-free Bioelectronic Detection.

Bioelectronic transducing surfaces that are nanometric in size have been the main route to detect single molecules. Though enabling the study of rarer events, such methodologies are not suited to assay at concentrations below the nanomolar level. Bioelectronic field-effect-transistors with a wide (µm2-mm2) transducing interface are also assumed to be not suited, because the molecule to be detected is orders of magnitude smaller than the transducing surface. Indeed, it is like seeing changes on the surface of a one-kilometer-wide pond when a droplet of water falls on it. However, it is a fact that a number of large-area transistors have been shown to detect at a limit of detection lower than femtomolar; they are also fast and hence innately suitable for point-of-care applications. This review critically discusses key elements, such as sensing materials, FET-structures, and target molecules that can be selectively assayed. The amplification effects enabling extremely sensitive large-area bioelectronic sensing are also addressed.

Thermal-Induced Performance Decay of the State-of-the-Art Polymer: Non-Fullerene Solar Cells and the Method of Suppression.

Improving thermal stability is of great importance for the industrialization of polymer solar cells (PSC). In this paper, we systematically investigated the high-temperature thermal annealing effect on the device performance of the state-of-the-art polymer:non-fullerene (PM6:Y6) solar cells with an inverted structure. Results revealed that the overall performance decay (19% decrease) was mainly due to the fast open-circuit voltage (VOC, 10% decrease) and fill factor (FF, 10% decrease) decays whereas short circuit current (JSC) was relatively stable upon annealing at 150 °C (0.5% decrease). Pre-annealing on the ZnO/PM6:Y6 at 150 °C before the completion of cell fabrication resulted in a 1.7% performance decrease, while annealing on the ZnO/PM6:Y6/MoO3 films led to a 10.5% performance decay, indicating that the degradation at the PM6:Y6/MoO3 interface is the main reason for the overall performance decay. The increased ideality factor and reduced built-in potential confirmed by dark J - V curve analysis further confirmed the increased interfacial charge recombination after thermal annealing. The interaction of PM6:Y6 and MoO3 was proved by UV-Vis absorption and XPS measurements. Such deep chemical doping of PM6:Y6 led to unfavorable band alignment at the interface, which led to increased surface charge recombination and reduced built-in potential of the cells after thermal annealing. Inserting a thin C60 layer between the PM6:Y6 and MoO3 significantly improved the cells' thermal stability, and less than 2% decay was measured for the optimized cell with 3 nm C60.

A large-area organic transistor with 3D-printed sensing gate for noninvasive single-molecule detection of pancreatic mucinous cyst markers.

Early diagnosis in a premalignant (or pre-invasive) state represents the only chance for cure in neoplastic diseases such as pancreatic-biliary cancer, which are otherwise detected at later stages and can only be treated using palliative approaches, with no hope for a cure. Screening methods for the purpose of secondary prevention are not yet available for these cancers. Current diagnostic methods mostly rely on imaging techniques and conventional cytopathology, but they do not display adequate sensitivity to allow valid early diagnosis. Next-generation sequencing can be used to detect DNA markers down to the physical limit; however, this assay requires labeling and is time-consuming. The additional determination of a protein marker that is a predictor of aggressive behavior is a promising innovative approach, which holds the potential to improve diagnostic accuracy. Moreover, the possibility to detect biomarkers in blood serum offers the advantage of a noninvasive diagnosis. In this study, both the DNA and protein markers of pancreatic mucinous cysts were analyzed in human blood serum down to the single-molecule limit using the SiMoT (single-molecule assay with a large transistor) platform. The SiMoT device proposed herein, which exploits an inkjet-printed organic semiconductor on plastic foil, comprises an innovative 3D-printed sensing gate module, consisting of a truncated cone that protrudes from a plastic substrate and is compatible with standard ELISA wells. This 3D gate concept adds tremendous control over the biosensing system stability, along with minimal consumption of the capturing molecules and body fluid samples. The 3D sensing gate modules were extensively characterized from both a material and electrical perspective, successfully proving their suitability as detection interfaces for biosensing applications. KRAS and MUC1 target molecules were successfully analyzed in diluted human blood serum with the 3D sensing gate functionalized with b-KRAS and anti-MUC1, achieving a limit of detection of 10 zM and 40 zM, respectively. These limits of detection correspond to (1 ± 1) KRAS and (2 ± 1) MUC1 molecules in the 100 µL serum sample volume. This study provides a promising application of the 3D SiMoT platform, potentially facilitating the timely, noninvasive, and reliable identification of pancreatic cancer precursor cysts.

Determination of Surface Recombination Velocities at Contacts in Organic Semiconductor Devices Using Injected Carrier Reservoirs.

A method to determine surface recombination velocities at collecting contacts in interface-limited organic semiconductor devices, based on the extraction of injected carrier reservoirs in a single-carrier sandwich-type structure, is presented. The analytical framework is derived and verified with drift-diffusion simulations. The method is demonstrated on solution-processed organic semiconductor devices with hole-blocking TiO_{2}/organic and SiO_{2}/organic interfaces, relevant for solar cell and transistor applications, respectively.

Environmentally friendly transistors and circuits on paper.

We created environmentally friendly low-voltage, ion-modulated transistors (IMTs) that can be fabricated successfully on a paper substrate. A range of ionic liquids (ILs) based on choline chloride (ChoCl) were used as the electrolytic layer in the IMTs. Different organic compounds were mixed with ChoCl to create solution-processable deep eutectic mixtures that are liquid or semiliquid at room temperature. In the final, solid version of the IMT, the ILs are also solidified by using a commercial binder to create printable transistor structures The semiconductor layer in the IMT is also substituted with a blend of the original semiconductor and a biodegradable polymer insulator. This reduces the amount of expensive and potentially harmful semiconductor used, and it also provides increased transistor performance, especially increasing the device switching speed. These environmentally friendly IMTs are then used to create ring oscillators, logic gates, and memories on paper.

Versatile characterization of thiol-functionalized printed metal electrodes on flexible substrates for cheap diagnostic applications.

BACKGROUND: Cheap, reliable, point-of-care diagnostics is a necessity for the growing and aging population of the world. Paper substrate and printing method, combined together, are the cheapest possible method for generating high-volume diagnostic sensor platforms. Electrical transduction tools also minimize the cost and enhance the simplicity of the devices. METHODS: Standard surface characterization techniques, namely contact angle measurements, atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) were used to analyze the growth of the organic thiol layers on top of the printed metal electrodes on paper substrates. The results were compared with those obtained by impedimetric electrical characterization method. RESULTS: This article reports the fabrication and characterization of printed metal electrodes and their functionalization by organic layers on paper and plastic substrates for biosensing and diagnostic applications. Impedimetric measurement is proposed as a simple, yet elegant, method of characterization of the organic layer growth. CONCLUSIONS: Very good correlation was observed between the results of organic layer growth from different measurement methods, justifying the use of paper as a substrate, printing as a method for fabricating metal and organic layers and impedance as a suitable measurement method for hand-held diagnostic devices. GENERAL SIGNIFICANCE: This result paves the way for the fabrication of more advanced bio-recognition layers for bio-affinity sensors using a printing technology that is compatible with flexible and cheap paper substrates. This article is part of a Special Issue entitled Organic Bioelectronics - Novel Applications in Biomedicine.

Impact of humidity on functionality of on-paper printed electronics.

A multilayer coated paper substrate, combining barrier and printability properties was manufactured utilizing a pilot-scale slide curtain coating technique. The coating structure consists of a thin mineral pigment layer coated on top of a barrier layer. The surface properties, i.e. smoothness and surface porosity, were adjusted by the choice of calendering parameters. The influence of surface properties on the fine line printability and conductivity of inkjet-printed silver lines was studied. Surface roughness played a significant role when printing narrow lines, increasing the risk of defects and discontinuities, whereas for wider lines the influence of surface roughness was less critical. A smooth, calendered surface resulted in finer line definition, i.e. less edge raggedness. Dimensional stability and its influence on substrate surface properties as well as on the functionality of conductive tracks and transistors were studied by exposure to high/low humidity cycles. The barrier layer of the multilayer coated paper reduced the dimensional changes and surface roughness increase caused by humidity and helped maintain the conductivity of the printed tracks. Functionality of a printed transistor during a short, one hour humidity cycle was maintained, but a longer exposure to humidity destroyed the non-encapsulated transistor.

An impedimetric study of DNA hybridization on paper-supported inkjet-printed gold electrodes.

In this study, two different supramolecular recognition architectures for impedimetric detection of DNA hybridization have been formed on disposable paper-supported inkjet-printed gold electrodes. The gold electrodes were fabricated using a gold nanoparticle based ink. The first recognition architecture consists of subsequent layers of biotinylated self-assembly monolayer (SAM), streptavidin and biotinylated DNA probe. The other recognition architecture is constructed by immobilization of thiol-functionalized DNA probe (HS-DNA) and subsequent backfill with 11-mercapto-1-undecanol (MUOH) SAM. The binding capacity and selectivity of the recognition architectures were examined by surface plasmon resonance (SPR) measurements. SPR results showed that the HS-DNA/MUOH system had a higher binding capacity for the complementary DNA target. Electrochemical impedance spectroscopy (EIS) measurements showed that the hybridization can be detected with impedimetric spectroscopy in picomol range for both systems. EIS signal indicated a good selectivity for both recognition architectures, whereas SPR showed very high unspecific binding for the HS-DNA/MUOH system. The factors affecting the impedance signal were interpreted in terms of the complexity of the supramolecular architecture. The more complex architecture acts as a less ideal capacitive sensor and the impedance signal is dominated by the resistive elements.

Industrial Kraft Lignin Based Binary Cathode Interface Layer Enables Enhanced Stability in High Efficiency Organic Solar Cells.

Herein, a binary cathode interface layer (CIL) strategy based on the industrial solvent fractionated LignoBoost kraft lignin (KL) is adopted for fabrication of organic solar cells (OSCs). The uniformly distributed phenol moieties in KL enable it to easily form hydrogen bonds with commonly used CIL materials, i.e., bathocuproine (BCP) and PFN-Br, resulting in binary CILs with tunable work function (WF). This work shows that the binary CILs work well in OSCs with large KL ratio compatibility, exhibiting equivalent or even higher efficiency to the traditional CILs in state of art OSCs. In addition, the combination of KL and BCP significantly enhanced OSC stability, owing to KL blocking the reaction between BCP and nonfullerene acceptors (NFAs). This work provides a simple and effective way to achieve high-efficient OSCs with better stability and sustainability by using wood-based materials.

Analysis of Clinical Samples of Pancreatic Cyst's Lesions with A Multi-Analyte Bioelectronic Simot Array Benchmarked Against Ultrasensitive Chemiluminescent Immunoassay.

Pancreatic cancer, ranking as the third factor in cancer-related deaths, necessitates enhanced diagnostic measures through early detection. In response, SiMoT-Single-molecule with a large Transistor multiplexing array, achieving a Technology Readiness Level of 5, is proposed for a timely identification of pancreatic cancer precursor cysts and is benchmarked against the commercially available chemiluminescent immunoassay SIMOA (Single molecule array) SP-X System. A cohort of 39 samples, comprising 33 cyst fluids and 6 blood plasma specimens, undergoes detailed examination with both technologies. The SiMoT array targets oncoproteins MUC1 and CD55, and oncogene KRAS, while the SIMOA SP-X planar technology exclusively focuses on MUC1 and CD55. Employing Principal Component Analysis (PCA) for multivariate data processing, the SiMoT array demonstrates effective discrimination of malignant/pre-invasive high-grade or potentially malignant low-grade pancreatic cysts from benign non-mucinous cysts. Conversely, PCA analysis applied to SIMOA assay reveals less effective differentiation ability among the three cyst classes. Notably, SiMoT unique capability of concurrently analyzing protein and genetic markers with the threshold of one single molecule in 0.1 mL positions it as a comprehensive and reliable diagnostic tool. The electronic response generated by the SiMoT array facilitates direct digital data communication, suggesting potential applications in the development of field-deployable liquid biopsy.

Experimental design of stencil-printed high-performance organic electrochemical transistors.

Organic electrochemical transistors (OECTs) are widely employed in several bioelectronic applications such as biosensors, logic circuits, and neuromorphic engineering, providing a seamless link between the realm of biology and electronics. More specifically, OECTs are endowed with remarkable signal amplification, the ability to operate in an aqueous environment, and the effective transduction of ionic to electrical signals. One main limiting factor preventing OECTs' wide use is the need for microfabrication processes, typically requiring specialized equipment. From this perspective, a robust and cost-effective production protocol to achieve high-performing OECT would be desirable. Herein, a straightforward stencil-printed OECT fabrication procedure is proposed, where the electrical performance can be controlled by adjusting the electronic channel fabrication conditions. An experimental design approach is undertaken to optimize OECT figures of merit by varying key parameters such as the annealing temperature and time, as well as the transistor active channel length. The resulting OECT devices, fabricated through a high-yield, cost-effective, and fast stencil printing technique, feature large transconductance values at low operating voltages. The experimental design allowed for minimizing the threshold voltage (VT = 260 mV) while keeping a high on/off ratio (7 × 103). A signal-to-noise ratio as high as 40 dB was obtained, which is among the highest for OECTs, operating in an aqueous electrolyte operated in a DC mode. An atomic force microscopy (AFM) characterization has been undertaken to analyze the channel morphology in the OECTs, correlating the annealing conditions with the charge transport properties.

Boosting Perovskite Solar Cells Efficiency and Stability: Interfacial Passivation of Crosslinked Fullerene Eliminates the "Burn-in" Decay.

Perovskite solar cells (PSCs) longevity is nowadays the bottleneck for their full commercial exploitation. Although lot of research is ongoing, the initial decay of the output power - an effect known as "burn-in" degradation happening in the first 100 h - is still unavoidable, significantly reducing the overall performance (typically of >20%). In this paper, the origin of the "burn-in" degradation in n-i-p type PSCs is demonstrated that is directly related to Li+ ions migration coming from the SnO2 electron transporting layer visualized by time-of-flight secondary ion mass spectrometry (TOF-SIMS) measurements. To block the ion movement, a thin cross-linked [6,6]-phenyl-C61-butyric acid methyl ester layer on top of the SnO2 layer is introduced, resulting in Li+ immobilization. This results in the elimination of the "burn-in" degradation, showing for the first time a zero "burn-in" loss in the performances while boosting device power conversion efficiency to >22% for triple-cation-based PSCs and >24% for formamidinium-based (FAPbI3 ) PSCs, proving the general validity of this approach and creating a new framework for the realization of stable PSCs devices.

A Single-Molecule Bioelectronic Portable Array for Early Diagnosis of Pancreatic Cancer Precursors.

A cohort of 47 patients is screened for pancreatic cancer precursors with a portable 96-well bioelectronic sensing-array for single-molecule assay in cysts fluid and blood plasma, deployable at point-of-care (POC). Pancreatic cancer precursors are mucinous cysts diagnosed with a sensitivity of at most 80% by state-of-the-art cytopathological molecular analyses (e.g., KRASmut DNA). Adding the simultaneous assay of proteins related to malignant transformation (e.g., MUC1 and CD55) is deemed essential to enhance diagnostic accuracy. The bioelectronic array proposed here, based on single-molecule-with-a-large-transistor (SiMoT) technology, can assay both nucleic acids and proteins at the single-molecule limit-of-identification (LOI) (1% of false-positives and false-negatives). It comprises an enzyme-linked immunosorbent assay (ELISA)-like 8 × 12-array organic-electronics disposable cartridge with an electrolyte-gated organic transistor sensor array, and a reusable reader, integrating a custom Si-IC chip, operating via software installed on a USB-connected smart device. The cartridge is complemented by a 3D-printed sensing gate cover plate. KRASmut , MUC1, and CD55 biomarkers either in plasma or cysts-fluid from 5 to 6 patients at a time, are multiplexed at single-molecule LOI in 1.5 h. The pancreatic cancer precursors are classified via a machine-learning analysis resulting in at least 96% diagnostic-sensitivity and 100% diagnostic-specificity. This preliminary study opens the way to POC liquid-biopsy-based early diagnosis of pancreatic-cancer precursors in plasma.

Effect of a large hole reservoir on the charge transport in TiO2/organic hybrid devices.

We have fabricated hybrid devices in the form of indium tin oxide/titanium dioxide/poly(3-hexylthiophene):[6,6]-phenyl C61 butyric acid methyl ester/copper (ITO/TiO(2)/P3HT:PCBM/Cu) to clarify the impact of the TiO(2)/P3HT:PCBM interface on the charge transport using the charge extraction by linearly increasing voltage (CELIV) technique. We found that a large equilibrium charge reservoir is accumulated at negative offsets at the TiO(2)/P3HT:PCBM interface leading to space charge limited extraction current (SCLC) transients. We show analytically the SCLC transient response and compare the experimental data to calculated SCLC at a linearly increasing voltage. The theoretical calculations indicate that the large charge reservoir at negative offset voltages is due to thermally generated charges combined with poor hole extraction at the ITO/TiO(2) contact, due to the hole blocking character of TiO(2).

Role of Surface Coverage and Film Quality of the TiO2 Electron Selective Layer for Optimal Hole-Blocking Properties.

Titanium dioxide (TiO2) is a commonly used electron selective layer in thin-film solar cells. The energy levels of TiO2 align well with those of most light-absorbing materials and facilitate extracting electrons while blocking the extraction of holes. In a device, this separates charge carriers and reduces recombination. In this study, we have evaluated the hole-blocking behavior of TiO2 compact layers using charge extraction by linearly increasing voltage in a metal-insulator-semiconductor structure (MIS-CELIV). This hole-blocking property was characterized as surface recombination velocity (S R) for holes at the interface between a semiconducting polymer and TiO2 layer. TiO2 layers of different thicknesses were prepared by sol-gel dip coating on two transparent conductive oxide substrates with different roughnesses. Surface coverage and film quality on both substrates were characterized using X-ray photoelectron spectroscopy and atomic force microscopy, along with its conductive imaging mode. Thicker TiO2 coatings provided better surface coverage, leading to reduced S R, unless the layers were otherwise defective. We found S R to be a more sensitive indicator of the overall film quality, as varying S R values were still observed among the films that looked similar in their characteristics via other methods.

Cross-Linking of Doped Organic Semiconductor Interlayers for Organic Solar Cells: Potential and Challenges.

Solution-processable interlayers are important building blocks for the commercialization of organic electronic devices such as organic solar cells. Here, the potential of cross-linking to provide an insoluble, stable, and versatile charge transport layer based on soluble organic semiconductors is studied. For this purpose, a photoreactive tris-azide cross-linker is synthesized. The capability of the small molecular cross-linker is illustrated by applying it to a p-doped polymer used as a hole transport layer in organic solar cells. High cross-linking efficiency and excellent charge extraction properties of the cross-linked doped hole transport layer are demonstrated. However, at high doping levels in the interlayer, the solar cell efficiency is found to deteriorate. Based on charge extraction measurements and numerical device simulations, it is shown that this is due to diffusion of dopants into the active layer of the solar cell. Thus, in the development of future cross-linker materials, care must be taken to ensure that they immobilize not only the host but also the dopants.

Surface Plasmon Resonance Assay for Label-Free and Selective Detection of HIV-1 p24 Protein.

The early detection of the human immunodeficiency virus (HIV) is of paramount importance to achieve efficient therapeutic treatment and limit the disease spreading. In this perspective, the assessment of biosensing assay for the HIV-1 p24 capsid protein plays a pivotal role in the timely and selective detection of HIV infections. In this study, multi-parameter-SPR has been used to develop a reliable and label-free detection method for HIV-1 p24 protein. Remarkably, both physical and chemical immobilization of mouse monoclonal antibodies against HIV-1 p24 on the SPR gold detecting surface have been characterized for the first time. The two immobilization techniques returned a capturing antibody surface coverage as high as (7.5 ± 0.3) × 1011 molecule/cm2 and (2.4 ± 0.6) × 1011 molecule/cm2, respectively. However, the covalent binding of the capturing antibodies through a mixed self-assembled monolayer (SAM) of alkanethiols led to a doubling of the p24 binding signal. Moreover, from the modeling of the dose-response curve, an equilibrium dissociation constant KD of 5.30 × 10-9 M was computed for the assay performed on the SAM modified surface compared to a much larger KD of 7.46 × 10-5 M extracted for the physisorbed antibodies. The chemically modified system was also characterized in terms of sensitivity and selectivity, reaching a limit of detection of (4.1 ± 0.5) nM and an unprecedented selectivity ratio of 0.02.

Organic Field-Effect Transistor Platform for Label-Free, Single-Molecule Detection of Genomic Biomarkers.

The increasing interest in technologies capable of tracking a biomarker down to the physical limit points toward new opportunities in early diagnostics of progressive diseases. Indeed, single-molecule detection technologies are foreseen to enable clinicians to associate the tiniest increase in a biomarker with the progression of a disease, particularly at its early stage. Bioelectronic organic transistors represent an extremely powerful tool to achieve label-free and single-molecule detection of clinically relevant biomarkers. These electronic devices are millimetric in size and in the future could be mass-produced at low cost. The core of the single molecule with a large transistor (SiMoT) platform, based on an electrolyte-gated field-effect transistor, is a gold gate electrode biofunctionalized with a self-assembled monolayer, a densely packed layer of recognition elements. So far, only the SiMoT detection of proteins, using the corresponding antibodies as recognition elements, has been reported. In this study, the SiMoT sensing response toward genomic biomarkers is proposed. Herein, the gate is functionalized with a genomic biomarker for multiple sclerosis (miR-182). This is relevant, not only because a limit of detection of a single molecule is achieved but also because it proves that the SiMoT label-free, single-molecule detection principle is the only one of its kind that can detect, by means of the same platform, both protein and genomic markers.

Investigation of Well-Defined Pinholes in TiO2 Electron Selective Layers Used in Planar Heterojunction Perovskite Solar Cells.

The recently introduced perovskite solar cell (PSC) technology is a promising candidate for providing low-cost energy for future demands. However, one major concern with the technology can be traced back to morphological defects in the electron selective layer (ESL), which deteriorates the solar cell performance. Pinholes in the ESL may lead to an increased surface recombination rate for holes, if the perovskite absorber layer is in contact with the fluorine-doped tin oxide (FTO) substrate via the pinholes. In this work, we used sol-gel-derived mesoporous TiO2 thin films prepared by block co-polymer templating in combination with dip coating as a model system for investigating the effect of ESL pinholes on the photovoltaic performance of planar heterojunction PSCs. We studied TiO2 films with different porosities and film thicknesses, and observed that the induced pinholes only had a minor impact on the device performance. This suggests that having narrow pinholes with a diameter of about 10 nm in the ESL is in fact not detrimental for the device performance and can even, to some extent improve their performance. A probable reason for this is that the narrow pores in the ordered structure do not allow the perovskite crystals to form interconnected pathways to the underlying FTO substrate. However, for ultrathin (~20 nm) porous layers, an incomplete ESL surface coverage of the FTO layer will further deteriorate the device performance.