Electrochemical Determination of Epinephrine in Pharmaceutical Preparation Using Laponite Clay-Modified Graphene Inkjet-Printed Electrode.

Epinephrine (EP, also called adrenaline) is a compound belonging to the catecholamine neurotransmitter family. It can cause neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, Huntington's disease and amyotrophic lateral sclerosis. This work describes an amperometric sensor for the electroanalytical detection of EP by using an inkjet-printed graphene electrode (IPGE) that has been chemically modified by a thin layer of a laponite (La) clay mineral. The ion exchange properties and permeability of the chemically modified electrode (denoted La/IPGE) were evaluated using multi-sweep cyclic voltammetry, while its charge transfer resistance was determined by electrochemical impedance spectroscopy. The results showed that La/IPGE exhibited higher sensitivity to EP compared to the bare IPGE. The developed sensor was directly applied for the determination of EP in aqueous solution using differential pulse voltammetry. Under optimized conditions, a linear calibration graph was obtained in the concentration range between 0.8 µM and 10 µM. The anodic peak current of EP was directly proportional to its concentration, leading to detection limits of 0.34 µM and 0.26 µM with bare IPGE and La/IPGE, respectively. The sensor was successfully applied for the determination of EP in pharmaceutical preparations. Recovery rates and the effects of interfering species on the detection of EP were evaluated to highlight the selectivity of the elaborated sensor.

A Rapidly Responsive Sensor for Wireless Detection of Early and Mature Microbial Biofilms.

Biofilm-associated infections, which are able to resist antibiotics, pose a significant challenge in clinical treatments. Such infections have been linked to various medical conditions, including chronic wounds and implant-associated infections, making them a major public-health concern. Early-detection of biofilm formation offers significant advantages in mitigating adverse effects caused by biofilms. In this work, we aim to explore the feasibility of employing a novel wireless sensor for tracking both early-stage and matured-biofilms formed by the medically relevant bacteria Staphylococcus aureus and Pseudomonas aeruginosa. The sensor utilizes electrochemical reduction of an AgCl layer bridging two silver legs made by inkjet-printing, forming a part of near-field-communication tag antenna. The antenna is interfaced with a carbon cloth designed to promote the growth of microorganisms, thereby serving as an electron source for reduction of the resistive AgCl into a highly-conductive Ag bridge. The AgCl-Ag transformation significantly alters the impedance of the antenna, facilitating wireless identification of an endpoint caused by microbial growth. To the best of our knowledge, this study for the first time presents the evidence showcasing that electrons released through the actions of bacteria can be harnessed to convert AgCl to Ag, thus enabling the wireless, battery-less, and chip-less early-detection of biofilm formation.

Adaptive slice-specific z-shimming for 2D spoiled gradient-echo sequences.

PURPOSE: To reduce the misbalance between compensation gradients and macroscopic field gradients, we introduce an adaptive slice-specific z-shimming approach for 2D spoiled multi-echo gradient-echoe sequences in combination with modeling of the signal decay. METHODS: Macroscopic field gradients were estimated for each slice from a fast prescan (15 seconds) and then used to calculate slice-specific compensation moments along the echo train. The coverage of the compensated field gradients was increased by applying three positive and three negative moments. With a forward model, which considered the effect of the slice profile, the z-shim moment, and the field gradient, R2∗ maps were estimated. The method was evaluated in phantom and in vivo measurements at 3 T and compared with a spoiled multi-echo gradient-echo and a global z-shimming approach without slice-specific compensation. RESULTS: The proposed method yielded higher SNR in R2∗ maps due to a broader range of compensated macroscopic field gradients compared with global z-shimming. In global white matter, the mean interquartile range, proxy for SNR, could be decreased to 3.06 s-1 with the proposed approach, compared with 3.37 s-1 for global z-shimming and 3.52 s-1 for uncompensated multi-echo gradient-echo. CONCLUSION: Adaptive slice-specific compensation gradients between echoes substantially improved the SNR of R2∗ maps, and the signal could also be rephased in anatomical areas, where it has already been completely dephased.

A new sensor based on an amino-montmorillonite-modified inkjet-printed graphene electrode for the voltammetric determination of gentisic acid.

An amperometric sensor based on an inkjet-printed graphene electrode (IPGE) modified with amine-functionalized montmorillonite (Mt-NH2) for the electroanalysis and quantification of gentisic acid (GA) has been developed. The organoclay used as IPGE modifier was prepared and characterized by infrared spectroscopy, X-ray diffraction, scanning electron microscopy, CHN elemental analysis, and thermogravimetry. The electrochemical features of the Mt-NH2/IPGE sensor were investigated by cyclic voltammetry and electrochemical impedance spectroscopy. The sensor exhibited charge selectivity ability which was exploited for the electrochemical oxidation of GA. The GA amperometric response was high in acidic medium (Brinton-Robinson buffer, pH 2) due to favorable interactions between the protonated amine groups and the negatively charged GA. Kinetic studies were also performed by cyclic voltammetry, and the obtained electron transfer rate constant of 11.3 s-1 indicated a fast direct electron transfer rate of GA to the electrode. An approach using differential pulse voltammetry was then developed for the determination of GA (at + 0.233 V vs. a pseudo Ag/Ag+ reference electrode), and under optimized conditions, the sensor showed high sensitivity, a wide working linear range from 1 to 21 µM (R2 = 0.999), and a low detection limit of 0.33 µM (0.051 ± 0.01 mg L-1). The proposed sensor was applied to quantify GA in a commercial red wine sample. The simple and rapid method developed using a cheap clay material could be employed for the determination of various phenolic acids.

Assessment and correction of macroscopic field variations in 2D spoiled gradient-echo sequences.

PURPOSE: To model and correct the dephasing effects in the gradient-echo signal for arbitrary RF excitation pulses with large flip angles in the presence of macroscopic field variations. METHODS: The dephasing of the spoiled 2D gradient-echo signal was modeled using a numerical solution of the Bloch equations to calculate the magnitude and phase of the transverse magnetization across the slice profile. Additionally, regional variations of the transmit RF field and slice profile scaling due to macroscopic field gradients were included. Simulations, phantom, and in vivo measurements at 3 T were conducted for R2∗ and myelin water fraction (MWF) mapping. RESULTS: The influence of macroscopic field gradients on R2∗ and myelin water fraction estimation can be substantially reduced by applying the proposed model. Moreover, it was shown that the dephasing over time for flip angles of 60° or greater also depends on the polarity of the slice-selection gradient because of phase variation along the slice profile. CONCLUSION: Substantial improvements in R2∗ accuracy and myelin water fraction mapping coverage can be achieved using the proposed model if higher flip angles are required. In this context, we demonstrated that the phase along the slice profile and the polarity of the slice-selection gradient are essential for proper modeling of the gradient-echo signal in the presence of macroscopic field variations.

The four-minute approach revisited: accelerating MRI-based multi-factorial age estimation.

OBJECTIVES: This feasibility study aimed to investigate the reliability of multi-factorial age estimation based on MR data of the hand, wisdom teeth and the clavicles with reduced acquisition time. METHODS: The raw MR data of 34 volunteers-acquired on a 3T system and using acquisition times (TA) of 3:46 min (hand), 5:29 min (clavicles) and 10:46 min (teeth)-were retrospectively undersampled applying the commercially available CAIPIRINHA technique. Automatic and radiological age estimation methods were applied to the original image data as well as undersampled data to investigate the reliability of age estimates with decreasing acquisition time. Reliability was investigated determining standard deviation (SSD) and mean (MSD) of signed differences, intra-class correlation (ICC) and by performing Bland-Altman analysis. RESULTS: Automatic age estimation generally showed very high reliability (SSD < 0.90 years) even for very short acquisition times (SSD ≈ 0.20 years for a total TA of 4 min). Radiological age estimation provided highly reliable results for images of the hand (ICC ≥ 0.96) and the teeth (ICC ≥ 0.79) for short acquisition times (TA = 16 s for the hand, TA = 2:21 min for the teeth), imaging data of the clavicles allowed for moderate acceleration (TA = 1:25 min, ICC ≥ 0.71). CONCLUSIONS: The results demonstrate that reliable multi-factorial age estimation based on MRI of the hand, wisdom teeth and the clavicles can be performed using images acquired with a total acquisition time of 4 min.

Tape-Stripping Electrochemical Detection of Melanoma.

A noninvasive electrochemical melanoma detection approach based on using adhesive tapes for collecting and fixing cells from a suspicious skin area and transferring the cells into a scanning electrochemical microscope (SECM) is presented. The adhesive layer collects the cells reproducibly and keeps them well adhered on the tape during experiments in an electrolyte solution. A melanoma biomarker, here the intracellular enzyme tyrosinase (TYR), was imaged on the tape-collected cells without further cell lysing using antibodies that were labeled with horseradish peroxidase (HRP). The HRP labels catalyzed the oxidation of a dissolved redox-active species, which was detected at a soft microelectrode, gently brushed in contact mode over the tape. The melanoma biomarker was first detected on tape-stripped samples with murine melanoma cells of different concentrations. Thereafter, increasing levels of TYR were recorded in cells that were collected from the skin of melanoma mouse models representing three different stages of tumor growth. Additionally, SECM results of tape-stripped different human melanoma cell lines were confirmed by previous studies based on traditionally fixed and permeabilized cells.

Inkjet-Printed Carbon Nanotube Electrodes for Measuring Pyocyanin and Uric Acid in a Wound Fluid Simulant and Culture Media.

Polyacrylamide-coated, carbon nanotube (PA/CNT) electrodes were prepared by an inkjet printing process and used to measure pyocyanin and uric acid in a wound fluid simulant at 37 °C. These two molecules are potential indicators of infection, and therefore their detection could prove useful for monitoring wound healing. Pyocyanin is a marker for the common wound bacterium Pseudomonas aeruginosa. Our long-term goal is to use these inexpensive and disposable electrodes to measure biomarkers of wound healing directly. In this proof-of-concept work, studies were performed in a wound fluid simulant to evaluate the stability of the electrodes and their responsiveness for the two bioanalytes. The PA/CNT inkjet-printed electrodes and electrical contacts were stable with unchanging physical and electrochemical properties in the wound fluid simulant over a 7-8-day period at 37 °C. The detection figures of merit for pyocyanin in the simulant at 37 °C were as follows: linear over the physiologically relevant range = 0.10 to 100 µmol L-1 (R2 = 0.9992), limit of detection = 0.10 µmol L-1 (S/N = 3), sensitivity = 35.6 ± 0.8 mA-L mol-1 and response variability ≤4% RSD. The detection figures of merit for uric acid in the simulant at 37 °C were as follows: linear over the physiologically relevant range = 100 to 1000 µmol L-1 (R2 = 0.9997), sensitivity = 2.83 ± 0.01 mA-L mol-1, and response variability ≤4% RSD. The limit of detection was not determined. The PA/CNT electrodes were also used to quantify pyocyanin concentrations in cell-free culture media from different strains of P. aeruginosa. The detected concentrations ranged from 1.00 ± 0.02 to 118 ± 6 µM depending on the strain.

Ultrafast 3D Bloch-Siegert B 1 + -mapping using variational modeling.

PURPOSE: Highly accelerated B 1 + -mapping based on the Bloch-Siegert shift to allow 3D acquisitions even within a brief period of a single breath-hold. THEORY AND METHODS: The B 1 + dependent Bloch-Siegert phase shift is measured within a highly subsampled 3D-volume and reconstructed using a two-step variational approach, exploiting the different spatial distribution of morphology and B 1 + -field. By appropriate variable substitution the basic non-convex optimization problem is transformed in a sequential solution of two convex optimization problems with a total generalized variation (TGV) regularization for the morphology part and a smoothness constraint for the B 1 + -field. The method is evaluated on 3D in vivo data with retro- and prospective subsampling. The reconstructed B 1 + -maps are compared to a zero-padded low resolution reconstruction and a fully sampled reference. RESULTS: The reconstructed B 1 + -field maps are in high accordance to the reference for all measurements with a mean error below 1% and a maximum of about 4% for acceleration factors up to 100. The minimal error for different sampling patterns was achieved by sampling a dense region in k-space center with acquisition times of around 10-12 s for 3D-acquistions. CONCLUSIONS: The proposed variational approach enables highly accelerated 3D acquisitions of Bloch-Siegert data and thus full liver coverage in a single breath hold.

Rapid T1 quantification from high resolution 3D data with model-based reconstruction.

PURPOSE: Magnetic resonance imaging protocols for the assessment of quantitative information suffer from long acquisition times since multiple measurements in a parametric dimension are required. To facilitate the clinical applicability, accelerating the acquisition is of high importance. To this end, we propose a model-based optimization framework in conjunction with undersampling 3D radial stack-of-stars data. THEORY AND METHODS: High resolution 3D T1 maps are generated from subsampled data by employing model-based reconstruction combined with a regularization functional, coupling information from the spatial and parametric dimension, to exploit redundancies in the acquired parameter encodings and across parameter maps. To cope with the resulting non-linear, non-differentiable optimization problem, we propose a solution strategy based on the iteratively regularized Gauss-Newton method. The importance of 3D-spectral regularization is demonstrated by a comparison to 2D-spectral regularized results. The algorithm is validated for the variable flip angle (VFA) and inversion recovery Look-Locker (IRLL) method on numerical simulated data, MRI phantoms, and in vivo data. RESULTS: Evaluation of the proposed method using numerical simulations and phantom scans shows excellent quantitative agreement and image quality. T1 maps from accelerated 3D in vivo measurements, e.g. 1.8 s/slice with the VFA method, are in high accordance with fully sampled reference reconstructions. CONCLUSIONS: The proposed algorithm is able to recover T1 maps with an isotropic resolution of 1 mm3 from highly undersampled radial data by exploiting structural similarities in the imaging volume and across parameter maps.

Point-of-care amperometric determination of L-dopa using an inkjet-printed carbon nanotube electrode modified with dandelion-like MnO2 microspheres.

An electrochemical sensor is described for the determination of L-dopa (levodopa; 3,4-dihydroxyphenylalanine). An inkjet-printed carbon nanotube (IJPCNT) electrode was modified with manganese dioxide microspheres by drop-casting. They coating was characterized by field emission scanning electron microscopy, Fourier-transform infrared spectroscopy and X-ray powder diffraction. The sensor, best operated at a working voltage of 0.3 V, has a linear response in the 0.1 to 10 µM L-dopa concentration range, a 54 nM detection limit, excellent reproducibility, repeatability and selectivity. The amperometric approach was applied to the determination of L-dopa in spiked biological fluids and displayed satisfactory accuracy and precision. Graphical abstract Schematic representation of an amperometric method for determination L-dopa. It is based on the use of inkjet-printed carbon nanotube electrode (IJPCNT) modified with manganese dioxide (MnO2).

Water-Repellent Low-Dimensional Fluorous Perovskite as Interfacial Coating for 20% Efficient Solar Cells.

Hybrid perovskite solar cells have been capturing an enormous research interest in the energy sector due to their extraordinary performances and ease of fabrication. However, low device lifetime, mainly due to material and device degradation upon water exposure, challenges their near-future commercialization. Here, we synthesized a new fluorous organic cation used as organic spacer to form a low-dimensional perovskite (LDP) with an enhanced water-resistant character. The LDP is integrated with three-dimensional (3D) perovskite absorbers in the form of MA0.9FA0.1PbI3 (FA = NH2CH = NH2+, MA = CH3NH3+) and Cs0.1FA0.74MA0.13PbI2.48Br0.39. In both cases, a LDP layer self-assembles as a thin capping layer on the top of the 3D bulk, making the perovskite surface hydrophobic. Our easy and robust approach, validated for different perovskite compositions, limits the interface deterioration in perovskite solar cells yielding to >20% power conversion efficient solar cells with improved stability, especially pronounced in the first hours of functioning under environmental conditions. As a consequence, single and multijunction perovskite devices, such as tandem solar cells, can benefit from the use of the waterproof stabilization here demonstrated, a concept which can be further expanded in the perovskite optoelectronic industry beyond photovoltaics.

Surface-Confined Electrochemiluminescence Microscopy of Cell Membranes.

Herein is reported a surface-confined microscopy based on electrochemiluminescence (ECL) that allows to image the plasma membrane of single cells at the interface with an electrode. By analyzing photoluminescence (PL), ECL and AFM images of mammalian CHO cells, we demonstrate that, in contrast to the wide-field fluorescence, ECL emission is confined to the immediate vicinity of the electrode surface and only the basal membrane of the cell becomes luminescent. The resulting ECL microscopy reveals details that are not resolved by classic fluorescence microscopy, without any light irradiation and specific setup. The thickness of the ECL-emitting regions is ∼500 nm due to the unique ECL mechanism that involves short-lifetime electrogenerated radicals. In addition, the reported ECL microscopy is a dynamic technique that reflects the transport properties through the cell membranes and not only the specific labeling of the membranes. Finally, disposable transparent carbon nanotube (CNT)-based electrodes inkjet-printed on classic microscope glass coverslips were used to image cells in both reflection and transmission configurations. Therefore, our approach opens new avenues for ECL as a surface-confined microscopy to develop single cell assays and to image the dynamics of biological entities in cells or in membranes.

Immuno-affinity Amperometric Detection of Bacterial Infections.

A combination of an immuno-affinity enrichment strategy and sensitive amperometric read-out was implemented in a point-of-care platform intended for bacterial infection analysis. Bacterial cells, selectively captured and enriched from complex matrices through immuno-affinity, were detected by amperometric monitoring of the redox state of metabolic activity indicators, providing species identification and viable-cell quantification. The method was successfully employed for the diagnosis of bacterial infections including antimicrobial susceptibility testing with only several hours of total working time.

Single Cell Electrochemiluminescence Imaging: From the Proof-of-Concept to Disposable Device-Based Analysis.

We report here the development of coreactant-based electrogenerated chemiluminescence (ECL) as a surface-confined microscopy to image single cells and their membrane proteins. Labeling the entire cell membrane allows one to demonstrate that, by contrast with fluorescence, ECL emission is only detected from fluorophores located in the immediate vicinity of the electrode surface (i.e., 1-2 µm). Then, to present the potential diagnostic applications of our approach, we selected carbon nanotubes (CNT)-based inkjet-printed disposable electrodes for the direct ECL imaging of a labeled plasma receptor overexpressed on tumor cells. The ECL fluorophore was linked to an antibody and enabled to localize the ECL generation on the cancer cell membrane in close proximity to the electrode surface. Such a result is intrinsically associated with the unique ECL mechanism and is rationalized by considering the limited lifetimes of the electrogenerated coreactant radicals. The electrochemical stimulus used for luminescence generation does not suffer from background signals, such as the typical autofluorescence of biological samples. The presented surface-confined ECL microscopy should find promising applications in ultrasensitive single cell imaging assays.

Soft Electrochemical Probes for Mapping the Distribution of Biomarkers and Injected Nanomaterials in Animal and Human Tissues.

Monitoring biomarkers and injected theranostic nanomaterials in tissues and organs plays a pivotal role in numerous medical applications ranging from cancer diagnostics to drug delivery. Scanning electrochemical microscopy has been demonstrated as a powerful tool to create highly resolved maps of the distributions of relevant biomolecules in cells and tissues without suffering from the optical interferences of conventional microscopy. We demonstrate for the first time the application of soft microelectrodes brushing in contact mode over large and thick tissues as well as organs that were immersed in an electrolyte solution. Amperometric currents were recorded based on the local flux of redox-active species locally and specifically generated by the biomarkers and nanomaterials to create maps of the biodistribution of graphene oxide nanoribbons in mouse livers, prognostic protein biomarkers in human melanoma and redox-active proteins in mouse heart.

Fixation and Permeabilization Approaches for Scanning Electrochemical Microscopy of Living Cells.

Scanning electrochemical microscopy (SECM) has been widely used for the electrochemical imaging of dynamic topographical and metabolic changes in alive adherent mammalian cells. However, extracting intracellular information by SECM is challenging, since it requires redox species to travel in and out the lipid cell membrane. Herein, we present cell fixation and permeabilization approaches as an alternative tool for visualizing cell properties by SECM. With this aim, adherent cells were analyzed in the SECM feedback mode in three different conditions: (i) alive; (ii) fixed, and (iii) fixed and permeabilized. The fixation was carried out with formaldehyde and does not damage lipid membranes. Therefore, this strategy can be used for the SECM investigation of cell topography or the passive transport of the redox mediator into the cells. Additional permeabilization of the cell membrane after fixation enables the analysis of the intracellular content through the coupling of SECM with immunoassay strategies for the detection of specific biomarkers. The latter was successfully applied as an easy and fast screening approach to detect the expression of the melanoma-associated marker tyrosinase in adherent melanoma cell lines corresponding to different cancer progression stages using the SECM substrate generation-tip collection mode. The present approach is simple, fast, and reliable and can open new ways to analyze cell cultures with electrochemically based scanning probe techniques.

Monitoring Tyrosinase Expression in Non-metastatic and Metastatic Melanoma Tissues by Scanning Electrochemical Microscopy.

Although tremendous progress has been made in the diagnosis of melanoma, the identification of different stages of malignancy in a reliable way remains challenging. Current strategies rely on optical monitoring of the concentration and spatial distribution of specific biomarkers. State-of-the-art optical methods can be affected by background-color interference and autofluorescence. We overcame these shortcomings by employing scanning electrochemical microscopy (SECM) to map the prognostic indicator tyrosinase (TyR) in non-metastatic and metastatic melanoma tissues by using soft-stylus microelectrodes. Electrochemical readout of the TyR distribution was enabled by adapting an immunochemical method. SECM can overcome the limitations of optical methods and opens unprecedented possibilities for improved diagnosis and understanding of the spatial distribution of TyR in different melanoma stages.

Inkjet printed nanohydrogel coated carbon nanotubes electrodes for matrix independent sensing.

Polyacrylamide (PA) based hydrogels are used in several applications including polyacrylamide gel electrophoresis and sensing devices. Homogeneous and compact PA films can be prepared based on chemical or photopolymerization processes. However, the accurate and reproducible coating of substrates with nanohydrogel patterns is challenging due to the in situ polymerization and deposition requirements. Herein, we report an inkjet printing (IJP) concept with simultaneously performed UV photopolymerization of a specifically prepared acrylamide/N,N'-methylenebis(acrylamide) containing ink. A prepolymerization step of the hydrogel precursor molecules was implemented in the ink formulation protocol to adjust the viscosity of the ink and to enhance the rate of polymerization during printing. After the optimization of the printing parameters, a nanometer thin PA hydrogel coating with well distributed nanopores was achieved on top of a stand-alone carbon nanotubes (CNTs) pattern. Batches of fully inkjet printed PA/CNT modified electrodes were prepared that showed outstanding improvements for the electrochemical detection of antioxidants in complex matrices such as untreated orange juice and red wine samples thanks to the properties of the PA coating.

Electrochemical push-pull probe: from scanning electrochemical microscopy to multimodal altering of cell microenvironment.

To understand biological processes at the cellular level, a general approach is to alter the cells' environment and to study their chemical responses. Herein, we present the implementation of an electrochemical push-pull probe, which combines a microfluidic system with a microelectrode, as a tool for locally altering the microenvironment of few adherent living cells by working in two different perturbation modes, namely electrochemical (i.e., electrochemical generation of a chemical effector compound) and microfluidic (i.e., infusion of a chemical effector compound from the pushing microchannel, while simultaneously aspirating it through the pulling channel, thereby focusing the flow between the channels). The effect of several parameters such as flow rate, working distance, and probe inclination angle on the affected area of adherently growing cells was investigated both theoretically and experimentally. As a proof of concept, localized fluorescent labeling and pH changes were purposely introduced to validate the probe as a tool for studying adherent cancer cells through the control over the chemical composition of the extracellular space with high spatiotemporal resolution. A very good agreement between experimental and simulated results showed that the electrochemical perturbation mode enables to affect precisely only a few living cells localized in a high-density cell culture.