Enhancing electrocatalytic activity in metallic thin films through surface segregation of carbon.

Thin layers of commonly used adhesion metals i.e., Cr and Ti were annealed to investigate and estimate their impact on the electrochemical properties of the carbon nanomaterials grown on top of them. The microstructure, surface chemistry, and electrochemical activities of these materials were evaluated and compared with those of as-deposited thin films. The results from X-ray photoelectron spectroscopy (XPS), Raman spectroscopy, grazing incidence X-ray diffraction (GIXRD), time-of-flight elastic recoil detection analysis (TOF-ERDA), and conductive atomic force microscopy (C-AFM) indicated the formation of a catalytic graphite layer on Cr following annealing, while no such layer was formed on Ti. This is attributed to the formation of the Cr2O3 layer on annealed Cr, which acts as a barrier to carbon diffusion into the underlying Cr. Conversely, Ti exhibits a high solubility for both carbon and oxygen, preventing the formation of the graphite layer. Cyclic voltammetry results showed that annealed Cr electrodes are electrochemically active towards both dopamine (DA) and ascorbic acid (AA) while no electrochemical activity is exhibited by annealed Ti. Quantum chemical calculations suggested that the presence of carbon as graphene or an amorphous form is critical for the oxidation reaction of probes. These results are significant for comprehending how the distinct solubilities of typical interstitial solutes influence the microstructure of adhesion metal layers and consequently yield diverse electrochemical properties.

Real-time selective detection of dopamine and serotonin at nanomolar concentration from complex in vitro systems.

Electrochemical sensors provide means for real-time monitoring of neurotransmitter release events, which is a relatively easy process in simple electrolytes. However, this does not apply to in vitro environments. In cell culture media, competitively adsorbing molecules are present at concentrations up to 350 000-fold excess compared to the neurotransmitter-of-interest. Because detection of dopamine and serotonin requires direct adsorption of the analyte to electrode surface, a significant loss of sensitivity occurs when recording is performed in the in vitro environment. Despite these challenges, our single-walled carbon nanotube (SWCNT) sensor was capable of selectively measuring dopamine and serotonin from cell culture medium at nanomolar concentration in real-time. A primary midbrain culture was used to prove excellent biocompatibility of our SWCNT electrodes, which is a necessity for brain-on-a-chip models. Most importantly, our sensor was able to electrochemically record spontaneous transient activity from dopaminergic cell culture without altering the culture conditions, which has not been possible earlier. Drug discovery and development requires high-throughput screening of in vitro models, being hindered by the challenges in non-invasive characterization of complex neuronal models such as organoids. Our neurotransmitter sensors could be used for real-time monitoring of complex neuronal models, providing an alternative tool for their characterization non-invasively.

Protein Adsorption and Its Effects on Electroanalytical Performance of Nanocellulose/Carbon Nanotube Composite Electrodes.

Protein fouling is a critical issue in the development of electrochemical sensors for medical applications, as it can significantly impact their sensitivity, stability, and reliability. Modifying planar electrodes with conductive nanomaterials that possess a high surface area, such as carbon nanotubes (CNTs), has been shown to significantly improve fouling resistance and sensitivity. However, the inherent hydrophobicity of CNTs and their poor dispersibility in solvents pose challenges in optimizing such electrode architectures for maximum sensitivity. Fortunately, nanocellulosic materials offer an efficient and sustainable approach to achieving effective functional and hybrid nanoscale architectures by enabling stable aqueous dispersions of carbon nanomaterials. Additionally, the inherent hygroscopicity and fouling-resistant nature of nanocellulosic materials can provide superior functionalities in such composites. In this study, we evaluate the fouling behavior of two nanocellulose (NC)/multiwalled carbon nanotube (MWCNT) composite electrode systems: one using sulfated cellulose nanofibers and another using sulfated cellulose nanocrystals. We compare these composites to commercial MWCNT electrodes without nanocellulose and analyze their behavior in physiologically relevant fouling environments of varying complexity using common outer- and inner-sphere redox probes. Additionally, we use quartz crystal microgravimetry with dissipation monitoring (QCM-D) to investigate the behavior of amorphous carbon surfaces and nanocellulosic materials in fouling environments. Our results demonstrate that the NC/MWCNT composite electrodes provide significant advantages for measurement reliability, sensitivity, and selectivity over only MWCNT-based electrodes, even in complex physiological monitoring environments such as human plasma.

Introduction of an electrochemical point-of-care assay for quantitative determination of paracetamol in finger-prick capillary whole blood samples.

AIMS: Measuring venous plasma paracetamol concentrations is time- and resource-consuming. We aimed to validate a novel electrochemical point-of-care (POC) assay for rapid paracetamol concentration determinations. METHODS: Twelve healthy volunteers received 1 g oral paracetamol, and its concentrations were analysed 10 times over 12 h for capillary whole blood (POC), venous plasma (high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS)), and dried capillary blood (HPLC-MS/MS). RESULTS: At concentrations >30 µM, POC showed upward biases of 20% (95% limits of agreement [LOA] -22 to 62) and 7% (95% LOA -23 to 38) compared with venous plasma and capillary blood HPLC-MS/MS, respectively. There were no significant differences between mean concentrations for the paracetamol elimination phase. CONCLUSIONS: Upward biases in POC compared with venous plasma HPLC-MS/MS were likely due to higher paracetamol concentrations in capillary blood than in venous plasma and to faulty individual sensors. The novel POC method is a promising tool for paracetamol concentration analysis.

Modulating the Geometry of the Carbon Nanofiber Electrodes Provides Control over Dopamine Sensor Performance.

One of the major challenges for in vivo electrochemical measurements of dopamine (DA) is to achieve selectivity in the presence of interferents, such as ascorbic acid (AA) and uric acid (UA). Complicated multimaterial structures and ill-defined pretreatments have been frequently utilized to enhance selectivity. The lack of control over the realized structures has prevented establishing associations between the achieved selectivity and the electrode structure. Owing to their easily tailorable structure, carbon nanofiber (CNF) electrodes have become promising materials for neurobiological applications. Here, a novel yet simple strategy to control the sensitivity and selectivity of CNF electrodes toward DA is reported. It consists of adjusting the lengths of CNF by modulating the growth phase during the fabrication process while keeping the surface chemistries similar. It was observed that the sensitivity of the CNF electrodes toward DA was enhanced with the increase in the fiber lengths. More importantly, the increase in the fiber length induced (i) an anodic shift in the DA oxidation peak and (ii) a cathodic shift in the AA oxidation peak. As the UA oxidation peak remained unaffected at high anodic potentials, the electrodes with long CNFs showed excellent selectivity. Electrodes without proper fibers showed only a single broad peak in the solution of AA, DA, and UA, completely lacking the ability to discriminate DA. Hence, the simple strategy of controlling CNF length without the need to carry out any complex chemical treatments provides us a feasible and robust route to fabricate electrode materials for neurotransmitter detection with excellent sensitivity and selectivity.

Ascorbic acid does not necessarily interfere with the electrochemical detection of dopamine.

It is widely stated that ascorbic acid (AA) interferes with the electrochemical detection of neurotransmitters, especially dopamine, because of their overlapping oxidation potentials on typical electrode materials. As the concentration of AA is several orders of magnitude higher than the concentration of neurotransmitters, detection of neurotransmitters is difficult in the presence of AA and requires either highly stable AA concentration or highly selective neurotransmitter sensors. In contrast to the common opinion, we show that AA does not always interfere electrochemical detection of neurotransmitters. The decay of AA is rapid in cell culture medium, having a half-time of 2.1 hours, according to which the concentration decreases by 93% in 8 hours and by 99.75% in 18 hours. Thus, AA is eventually no longer detected by electrodes and the concentration of neurotransmitters can be effectively monitored. To validate this claim, we used unmodified single-wall carbon nanotube electrode to measure dopamine at physiologically relevant concentration range (25-1000 nM) from human midbrain organoid medium with highly linear response. Finally, AA is known to affect dopamine oxidation current through regeneration of dopamine, which complicates precise detection of small amounts of dopamine. By designing experiments as described here, this complication can be completely eliminated.

Accurate Computational Prediction of Core-Electron Binding Energies in Carbon-Based Materials: A Machine-Learning Model Combining Density-Functional Theory and GW.

We present a quantitatively accurate machine-learning (ML) model for the computational prediction of core-electron binding energies, from which X-ray photoelectron spectroscopy (XPS) spectra can be readily obtained. Our model combines density functional theory (DFT) with GW and uses kernel ridge regression for the ML predictions. We apply the new approach to disordered materials and small molecules containing carbon, hydrogen, and oxygen and obtain qualitative and quantitative agreement with experiment, resolving spectral features within 0.1 eV of reference experimental spectra. The method only requires the user to provide a structural model for the material under study to obtain an XPS prediction within seconds. Our new tool is freely available online through the XPS Prediction Server.

Deriving stair-climbing performance outcome measures using the smartphone barometer: Results of an algorithm development study.

As we seek to gain richer insights to understand intervention effects, and increasingly decentralise aspects of clinical trials to simplify participation, there is a growing interest in leveraging wearables and sensors to generate novel and informative clinical outcome measures for at-home assessment. The sensors embedded within smartphone technology provide one approach to capture of this data, and may be particularly useful when patients are already using mobile devices for at-home capture of other clinical trials data, such as patient-reported outcomes. We describe the results of an initial algorithm development study to determine whether the atmospheric pressure data provided by an onboard smartphone sensor is sufficiently informative to enable detection of a small height gain, such as that achieved during a short stair climb performance test. We were able to sufficiently distinguish height changes of 0.6 m in indoor conditions, representing around 4 stairs on an average staircase. This suggests that the smartphone barometer may indeed be suitable for inclusion within future work developing a stair-climbing performance outcome test instrumented using a mobile application.

Nanoscale geometry determines mechanical biocompatibility of vertically aligned nanofibers.

Vertically aligned carbon nanofibers (VACNFs) are promising material candidates for neural biosensors due to their ability to detect neurotransmitters in physiological concentrations. However, the expected high rigidity of CNFs could induce mechanical mismatch with the brain tissue, eliciting formation of a glial scar around the electrode and thus loss of functionality. We have evaluated mechanical biocompatibility of VACNFs by growing nickel-catalyzed carbon nanofibers of different lengths and inter-fiber distances. Long nanofibers with large inter-fiber distance prevented maturation of focal adhesions, thus constraining cells from obtaining a highly spread morphology that is observed when astrocytes are being contacted with stiff materials commonly used in neural implants. A silicon nanopillar array with 500 nm inter-pillar distance was used to reveal that this inhibition of focal adhesion maturation occurs due to the surface nanoscale geometry, more precisely the inter-fiber distance. Live cell atomic force microscopy was used to confirm astrocytes being significantly softer on the long Ni-CNFs compared to other surfaces, including a soft gelatin hydrogel. We also observed hippocampal neurons to mature and form synaptic contacts when being cultured on both long and short carbon nanofibers, without having to use any adhesive proteins or a glial monoculture, indicating high cytocompatibility of the material also with neuronal population. In contrast, neurons cultured on a planar tetrahedral amorphous carbon sample showed immature neurites and indications of early-stage apoptosis. Our results demonstrate that mechanical biocompatibility of biomaterials is greatly affected by their nanoscale surface geometry, which provides means for controlling how the materials and their mechanical properties are perceived by the cells. STATEMENT OF SIGNIFICANCE: Our research article shows, how nanoscale surface geometry determines mechanical biocompatibility of apparently stiff materials. Specifically, astrocytes were prevented from obtaining highly spread morphology when their adhesion site maturation was inhibited, showing similar morphology on nominally stiff vertically aligned carbon fiber (VACNF) substrates as when being cultured on ultrasoft surfaces. Furthermore, hippocampal neurons matured well and formed synapses on these carbon nanofibers, indicating high biocompatibility of the materials. Interestingly, the same VACNF materials that were used in this study have earlier also been proven to be capable for electrophysiological recordings and sensing neurotransmitters at physiological concentrations with ultra-high sensitivity and selectivity, thus providing a platform for future neural probes or smart culturing surfaces with superior sensing performance and biocompatibility.

Nanostructured Geometries Strongly Affect Fouling of Carbon Electrodes.

Electrode fouling is a major factor that compromises the performance of biosensors in in vivo usage. It can be roughly classified into (i) electrochemical fouling, caused by the analyte and its reaction products, and (ii) biofouling, caused by proteins and other species in the measurement environment. Here, we examined the effect of electrochemical fouling [in phosphate buffer saline (PBS)], biofouling [in cell-culture media (F12-K) with and without proteins], and their combination on the redox reactions occurring on carbon-based electrodes possessing distinct morphologies and surface chemistry. The effect of biofouling on the electrochemistry of an outer sphere redox probe, [Ru(NH3)6]3+, was negligible. On the other hand, fouling had a marked effect on the electrochemistry of an inner sphere redox probe, dopamine (DA). We observed that the surface geometry played a major role in the extent of fouling. The effect of biofouling on DA electrochemistry was the worst on planar pyrolytic carbon, whereas the multiwalled carbon nanotube/tetrahedral amorphous carbon (MWCNT/ta-C), possessing spaghetti-like morphology, and carbon nanofiber (CNF/ta-C) electrodes were much less seriously affected. The blockage of the adsorption sites for DA by proteins and other components of biological media and electrochemical fouling components (byproducts of DA oxidation) caused rapid surface poisoning. PBS washing for 10 consecutive cycles at 50 mV/s did not improve the electrode performance, except for CNF/ta-C, which performed better after PBS washing. Overall, this study emphasizes the combined effect of biological and electrochemical fouling to be critical for the evaluation of the functionality of a sensor. Thus, electrodes possessing composite nanostructures showed less surface fouling in comparison to those possessing planar geometry.

X-ray Spectroscopy Fingerprints of Pristine and Functionalized Graphene.

In this work, we demonstrate how to identify and characterize the atomic structure of pristine and functionalized graphene materials from a combination of computational simulation of X-ray spectra, on the one hand, and computer-aided interpretation of experimental spectra, on the other. Despite the enormous scientific and industrial interest, the precise structure of these 2D materials remains under debate. As we show in this study, a wide range of model structures from pristine to heavily oxidized graphene can be studied and understood with the same approach. We move systematically from pristine to highly oxidized and defective computational models, and we compare the simulation results with experimental data. Comparison with experiments is valuable also the other way around; this method allows us to verify that the simulated models are close to the real samples, which in turn makes simulated structures amenable to several computational experiments. Our results provide ab initio semiquantitative information and a new platform for extended insight into the structure and chemical composition of graphene-based materials.

Connection between the physicochemical characteristics of amorphous carbon thin films and their electrochemical properties.

Connecting a material's surface chemistry with its electrocatalytic performance is one of the major questions in analytical electrochemistry. This is especially important in many sensor applications where analytes from complex media need to be measured. Unfortunately, today this connection is still largely missing except perhaps for the most simple ideal model systems. Here we present an approach that can be used to obtain insights about this missing connection and apply it to the case of carbon nanomaterials. In this paper we show that by combining advanced computational techniques augmented by machine learning methods with x-ray absorption spectroscopy (XAS) and electrochemical measurements, it is possible to obtain a deeper understanding of the correlation between local surface chemistry and electrochemical performance. As a test case we show how by computationally assessing the growth of amorphous carbon (a-C) thin films at the atomic level, we can create computational structural motifs that may in turn be used to deconvolute the XAS data from the real samples resulting in local chemical information. Then, by carrying out electrochemical measurements on the same samples from which x-ray spectra were measured and that were further characterized computationally, it is possible to gain insight into the interplay between the local surface chemistry and electrochemical performance. To demonstrate this methodology, we proceed as follows: after assessing the basic electrochemical properties of a-C films, we investigate the effect of short HNO3treatment on the sensitivity of these electrodes towards an inner sphere redox probe dopamine to gain knowledge about the influence of altered surface chemistry to observed electrochemical performance. These results pave the way towards a more general assessment of electrocatalysis in different systems and provide the first steps towards data driven tailoring of electrode surfaces to gain optimal performance in a given application.

Electrochemical Detection of Morphine in Untreated Human Capillary Whole Blood.

Disposable single-use electrochemical sensor strips were used for quantitative detection of small concentrations of morphine in untreated capillary whole blood. Single-walled carbon nanotube (SWCNT) networks were fabricated on a polymer substrate to produce flexible, reproducible sensor strips with integrated reference and counter electrodes, compatible with industrial-scale processes. A thin Nafion coating was used on top of the sensors to enable direct electrochemical detection in whole blood. These sensors were shown to detect clinically relevant concentrations of morphine both in buffer and in whole blood samples. Small 38 µL finger-prick blood samples were spiked with 2 µL of morphine solution of several concentrations and measured without precipitation of proteins or any other further pretreatment. A linear range of 0.5-10 µM was achieved in both matrices and a detection limit of 0.48 µM in buffer. In addition, to demonstrate the applicability of the sensor in a point-of-care device, single-determination measurements were done with capillary samples from three subjects. An average recovery of 60% was found, suggesting that the sensor only measures the free, unbound fraction of the drug. An interference study with other opioids and possible interferents showed the selectivity of the sensor. This study clearly indicates that these Nafion/SWCNT sensor strips show great promise as a point-of-care rapid test for morphine in blood.

Disposable Nafion-Coated Single-Walled Carbon Nanotube Test Strip for Electrochemical Quantitative Determination of Acetaminophen in a Finger-Prick Whole Blood Sample.

A disposable electrochemical test strip for the quantitative point-of-care (POC) determination of acetaminophen (paracetamol) in plasma and finger-prick whole blood was fabricated. The industrially scalable dry transfer process of single-walled carbon nanotubes (SWCNTs) and screen printing of silver were combined to produce integrated electrochemical test strips. Nafion coating stabilized the potential of the Ag reference electrode and enabled the selective detection in spiked plasma as well as in whole blood samples. The test strips were able to detect acetaminophen in small 40 µL samples with a detection limit of 0.8 µM and a wide linear range from 1 µM to 2 mM, well within the required clinical range. After a simple 1:1 dilution of plasma and whole blood, a quantitative detection with good recoveries of 79% in plasma and 74% in whole blood was achieved. These results strongly indicate that these electrodes can be used directly to determine the unbound acetaminophen fraction without the need for any additional steps. The developed test strip shows promise as a rapid and simple POC quantitative acetaminophen assay.

Biofouling affects the redox kinetics of outer and inner sphere probes on carbon surfaces drastically differently - implications to biosensing.

Biofouling imposes a significant threat for sensing probes used in vivo. Antifouling strategies commonly utilize a protective layer on top of the electrode but this may compromise performance of the electrode. Here, we investigated the effect of surface topography and chemistry on fouling without additional protective layers. We have utilized two different carbon materials; tetrahedral amorphous carbon (ta-C) and SU-8 based pyrolytic carbon (PyC) in their typical smooth thin film structure as well as with a nanopillar topography templated from black silicon. The near edge X-ray absorption fine structure (NEXAFS) spectrum revealed striking differences in chemical functionalities of the surfaces. PyC contained equal amounts of ketone, hydroxyl and ether/epoxide groups, while ta-C contained significant amounts of carbonyl groups. Overall, oxygen functionalities were significantly increased on nanograss surfaces compared to the flat counterparts. Neither bovine serum albumin (BSA) or fetal bovine serum (FBS) fouling caused major effects on electron transfer kinetics of outer sphere redox (OSR) probe Ru(NH3)63+ on any of the materials. In contrast, negatively charged OSR probe IrCl62- kinetics were clearly affected by fouling, possibly due to the electrostatic repulsion between redox species and the anionically-charged proteins adsorbed on the electrode and/or stronger interaction of the proteins and positively charged surface. The OSR probe kinetics were less affected by fouling on PyC, probably due to conformational changes of proteins on the surface. Dopamine (DA) was tested as an inner sphere redox (ISR) probe and as expected, the kinetics were heavily dependent on the material; PyC had very fast electron transfer kinetics, while ta-C had sluggish kinetics. DA electron transfer kinetics were heavily affected on all surfaces by fouling (ΔEp increase 30-451%). The effect was stronger on PyC, possibly due to the more strongly adhered protein layer limiting the access of the probe to the inner sphere.

Electrochemical Detection of Oxycodone and Its Main Metabolites with Nafion-Coated Single-Walled Carbon Nanotube Electrodes.

Oxycodone is a strong opioid frequently used as an analgesic. Although proven efficacious in the management of moderate to severe acute pain and cancer pain, use of oxycodone imposes a risk of adverse effects such as addiction, overdose, and death. Fast and accurate determination of oxycodone blood concentration would enable personalized dosing and monitoring of the analgesic as well as quick diagnostics of possible overdose in emergency care. However, in addition to the parent drug, several metabolites are always present in the blood after a dose of oxycodone, and to date, there is no electrochemical data available on any of these metabolites. In this paper, a single-walled carbon nanotube (SWCNT) electrode and a Nafion-coated SWCNT electrode were used, for the first time, to study the electrochemical behavior of oxycodone and its two main metabolites, noroxycodone and oxymorphone. Both electrode types could selectively detect oxycodone in the presence of noroxycodone and oxymorphone. However, we have previously shown that addition of a Nafion coating on top of the SWCNT electrode is essential for direct measurements in complex biological matrices. Thus, the Nafion/SWCNT electrode was further characterized and used for measuring clinically relevant concentrations of oxycodone in buffer solution. The limit of detection for oxycodone with the Nafion/SWCNT sensor was 85 nM, and the linear range was 0.5-10 µM in buffer solution. This study shows that the fabricated Nafion/SWCNT sensor has potential to be applied in clinical concentration measurements.

Integrating Carbon Nanomaterials with Metals for Bio-sensing Applications.

Age structure in most developed countries is changing fast as the average lifespan is increasing significantly, calling for solutions to provide improved treatments for age-related neurological diseases and disorders. In order to address these problems, a reliable way of recording information about neurotransmitters from in vitro and in vivo applications is needed to better understand neurological diseases and disorders as well as currently used treatments. Likewise, recent developments in medicine, especially with the opioid crisis, are demanding a swift move to personalized medicine to administer patient needs rather than population-wide averages. In order to enable the so-called personalized medicine, it is necessary to be able to do measurements in vivo and in real time. These actions require sensitive and selective detection of different analytes from very demanding environments. Current state-of-the-art materials are unable to provide sensitive and selective detection of neurotransmitters as well as the required time resolution needed for drug molecules at a reasonable cost. To meet these challenges, we have utilized different metals to grow carbon nanomaterials and applied them for sensing applications showing that there are clear differences in their electrochemical properties based on the selected catalyst metal. Additionally, we have combined atomistic simulations to support optimizing materials for experiments and to gain further understanding of the atomistic level reactions between different analytes and the sensor surface. With carbon nanostructures grown from Ni and Al + Co + Fe hybrid, we can detect dopamine, ascorbic acid, and uric acid simultaneously. On the other hand, nanostructures grown from platinum provide a feasible platform for detection of H2O2 making them suitable candidates for enzymatic biosensors for detection of glutamate, for example. Tetrahedral amorphous carbon electrodes have an ability to detect morphine, paracetamol, tramadol, and O-desmethyltramadol. With carbon nanomaterial-based sensors, it is possible to reach metal-like properties in sensing applications using only a fraction of the metal as seed for the material growth. We have also seen that by using nanodiamonds as growth catalyst for carbon nanofibers, it is not possible to detect dopamine and ascorbic acid simultaneously, although the morphology of the resulting nanofibers is similar to the ones grown using Ni. This further indicates the importance of the metal selection for specific applications. However, Ni as a continuous layer or as separate islands does not provide adequate performance. Thus, it appears that metal nanoparticles combined with fiber-like morphology are needed for optimized sensor performance for neurotransmitter detection. This opens up a new research approach of application-specific nanomaterials, where carefully selected metals are integrated with carbon nanomaterials to match the needs of the sensing application in question.

Simultaneous Detection of Morphine and Codeine in the Presence of Ascorbic Acid and Uric Acid and in Human Plasma at Nafion Single-Walled Carbon Nanotube Thin-Film Electrode.

In clinical settings, the dosing and differential diagnosis of the poisoning of morphine (MO) and codeine (CO) is challenging due to interindividual variations in metabolism. However, direct electrochemical detection of these analytes from biological matrices is inherently challenging due to interference from large concentrations of anions, such as ascorbic acid (AA) and uric acid (UA), as well as fouling of the electrode by proteins. In this work, a disposable Nafion-coated single-walled carbon nanotube network (SWCNT) electrode was developed. We show facile electron transfer and efficient charge separation between the interfering anions and positively charged MO and CO, as well as significantly reduced matrix effect in human plasma. The Nafion coating alters the voltammetric response of MO and CO, enabling simultaneous detection. With this SWCNT/Nafion electrode, two linear ranges of 0.05-1 and 1-10 µM were found for MO and one linear range of 0.1-50 µM for CO. Moreover, the selective and simultaneous detection of MO and CO was achieved in large excess of AA and UA, as well as, for the first time, in unprocessed human plasma. The favorable properties of this electrode enabled measurements in plasma with only mild dilution and without the precipitation of proteins.

Fabrication of Micro- and Nanopillars from Pyrolytic Carbon and Tetrahedral Amorphous Carbon.

Pattern formation of pyrolyzed carbon (PyC) and tetrahedral amorphous carbon (ta-C) thin films were investigated at micro- and nanoscale. Micro- and nanopillars were fabricated from both materials, and their biocompatibility was studied with cell viability tests. Carbon materials are known to be very challenging to pattern. Here we demonstrate two approaches to create biocompatible carbon features. The microtopographies were 2 µ m or 20 µ m pillars (1:1 aspect ratio) with three different pillar layouts (square-grid, hexa-grid, or random-grid orientation). The nanoscale topography consisted of random nanopillars fabricated by maskless anisotropic etching. The PyC structures were fabricated with photolithography and embossing techniques in SU-8 photopolymer which was pyrolyzed in an inert atmosphere. The ta-C is a thin film coating, and the structures for it were fabricated on silicon substrates. Despite different fabrication methods, both materials were formed into comparable micro- and nanostructures. Mouse neural stem cells were cultured on the samples (without any coatings) and their viability was evaluated with colorimetric viability assay. All samples expressed good biocompatibility, but the topography has only a minor effect on viability. Two µ m pillars in ta-C shows increased cell count and aggregation compared to planar ta-C reference sample. The presented materials and fabrication techniques are well suited for applications that require carbon chemistry and benefit from large surface area and topography, such as electrophysiological and -chemical sensors for in vivo and in vitro measurements.