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1.  Microfabricated Microelectrode Sensor for Measuring Background and Slowly Changing Dopamine Concentrations 
The electrochemical detection of neurotransmitters in vivo has centered on fast scan cyclic voltammetry (FSCV) due to its temporal resolution, sensitivity and chemical selectivity. FSCV is a differential technique that records phasic (second-to-second) changes in the concentration of electroactive neurotransmitters such as dopamine (DA). To isolate the currents due to fluctuations in analyte concentration, in other words to make these phasic measurements, requires the subtraction of a large background current. The subtraction of this background and its volatility renders FSCV unable to determine background or slowly varying concentrations of electroactive analytes. However, there is still a need to readily determine the background and slowly changing concentrations of electroactive analytes in tissue. For example, the background concentrations of DA vary throughout the brain and can affect the dynamics of dopaminergic systems. So, this report presents a microfabricated electrochemical sensor for measuring background and slowly changing concentrations of DA in vitro with the selectivity and sensitivity of FSCV. The sensor is comprised of two microfabricated microelectrodes which are spaced 8 μm apart. Varying the applied potential of the outer electrode manipulates the local concentration of electroactive species including concentration at the inner electrode. These changes are measured at the inner electrode using FSCV. The resulting signal with calibration can determine the background and slowly changing concentration of DA with the selectivity and sensitivity of FSCV. In this study the background of DA is determined in vitro using this sensor. The DA signal is shown to be the result of adsorption/desorption at the outer electrode. Interference from ascorbate on the DA signal is shown to be minimal for this approach.
doi:10.1016/j.jelechem.2013.01.022
PMCID: PMC3607536  PMID: 23539293
2.  Evaluation of Three Field-Based Methods for Quantifying Soil Carbon 
PLoS ONE  2013;8(1):e55560.
Three advanced technologies to measure soil carbon (C) density (g C m−2) are deployed in the field and the results compared against those obtained by the dry combustion (DC) method. The advanced methods are: a) Laser Induced Breakdown Spectroscopy (LIBS), b) Diffuse Reflectance Fourier Transform Infrared Spectroscopy (DRIFTS), and c) Inelastic Neutron Scattering (INS). The measurements and soil samples were acquired at Beltsville, MD, USA and at Centro International para el Mejoramiento del Maíz y el Trigo (CIMMYT) at El Batán, Mexico. At Beltsville, soil samples were extracted at three depth intervals (0–5, 5–15, and 15–30 cm) and processed for analysis in the field with the LIBS and DRIFTS instruments. The INS instrument determined soil C density to a depth of 30 cm via scanning and stationary measurements. Subsequently, soil core samples were analyzed in the laboratory for soil bulk density (kg m−3), C concentration (g kg−1) by DC, and results reported as soil C density (kg m−2). Results from each technique were derived independently and contributed to a blind test against results from the reference (DC) method. A similar procedure was employed at CIMMYT in Mexico employing but only with the LIBS and DRIFTS instruments. Following conversion to common units, we found that the LIBS, DRIFTS, and INS results can be compared directly with those obtained by the DC method. The first two methods and the standard DC require soil sampling and need soil bulk density information to convert soil C concentrations to soil C densities while the INS method does not require soil sampling. We conclude that, in comparison with the DC method, the three instruments (a) showed acceptable performances although further work is needed to improve calibration techniques and (b) demonstrated their portability and their capacity to perform under field conditions.
doi:10.1371/journal.pone.0055560
PMCID: PMC3561178  PMID: 23383225
3.  Characterization of Local pH Changes in Brain Using Fast-Scan Cyclic Voltammetry with Carbon Microelectrodes 
Analytical chemistry  2010;82(23):9892-9900.
Transient local pH changes in the brain are important markers of neural activity that can be used to follow metabolic processes that underlie the biological basis of behavior, learning and memory. There are few methods that can measure pH fluctuations with sufficient time resolution in freely moving animals. Previously, fast-scan cyclic voltammetry at carbon-fiber microelectrodes was used for the measurement of such pH transients. However, the origin of the potential dependent current in the cyclic voltammograms for pH changes recorded in vivo was unclear. The current work explored the nature of these peaks and established the origin for some of them. A peak relating to the capacitive nature of the pH CV was identified. Adsorption of electrochemically inert species, such as aromatic amines and calcium could suppress this peak, and is the origin for inconsistencies regarding in vivo and in vitro data. Also, we identified an extra peak in the in vivo pH CV relating to the presence of 3,4-dihydroxyacetic acid (DOPAC) in the brain extracellular fluid. To evaluate the in vivo performance of the carbon-fiber sensor, carbon dioxide inhalation by an anesthetized rat was used to induce brain acidosis induced by hypercapnia. Hypercapnia is demonstrated to be a useful tool to induce robust in vivo pH changes, allowing confirmation of the pH signal observed with FSCV.
doi:10.1021/ac102399n
PMCID: PMC2995839  PMID: 21047096
pH sensor; carbon-fiber microelectrode; in vivo voltammetry; hypercapnia; acidosis; adsorption; FSCV
4.  Microfabricated FSCV-Compatible Microelectrode Array for Real-time Monitoring of Heterogeneous Dopamine Release 
The Analyst  2010;135(7):1556-1563.
Fast scan cyclic voltammetry (FSCV) has been used previously to detect neurotransmitter release and reuptake in vivo. An advantage that FSCV has over other electrochemical techniques is its ability to distinguish neurotransmitters of interest (i.e. monoamines) from their metabolites using their respective characteristic cyclic voltammogram. While much has been learned with this technique, it has generally only been used in a single working electrode arrangement. Additionally, traditional electrode fabrication techniques tend to be difficult and somewhat irreproducible. Described in this report is a fabrication method for a FSCV compatible microelectrode array (FSCV-MEA) that is capable of functioning in vivo. The microfabrication techniques employed here allow for better reproducibility than traditional fabrication methods of carbon fiber microelectrodes, and enable batch fabrication of electrode arrays. The reproducibility and electrochemical qualities of the probes were assessed along with cross talk in vitro. Heterogeneous release of electrically stimulated dopamine was observed in real-time in the striatum of an anesthetized rat using the FSCV-MEA. The heterogeneous effects of pharmacology on the striatum was also observed and shown to be consistent across multiple animals.
doi:10.1039/c0an00114g
PMCID: PMC2975426  PMID: 20464031
5.  Enhancing Electrochemical Detection by Scaling Solid State Nanogaps 
The ability to quickly and inexpensively fabricate planar solid state nanogaps has enabled research to be effectively performed on devices down to just a few nanometers. Here, nanofabricated electrode pairs with electrode-to-electrode spacings of <4, 6 and 20 nm are utilized for monitoring an electroactive molecules, dopamine, in ionic solution. The results show a several order of magnitude enhancement of the electrochemical signal, collected current, for the solid state nanogaps with 6 nm electrode-electrode spacings as compared to traditional microelectrodes. The data from the <4 nm and 20 nm solid state nanogaps verify that this enhancement is due to cycling of the redox molecules in the confined geometry of the nanogap. In addition the data collected for the <4 nm nanogap emphasizes and reinforces that scaling does have limits and that as device sizes move to the few nanometer scale, the influence of a molecule's size and other physical properties becomes increasingly important and can eventually dominate the generated signals.
doi:10.1016/j.jelechem.2010.03.018
PMCID: PMC2863020  PMID: 20454636
6.  Carbon Microelectrodes with a Renewable Surface 
Analytical chemistry  2010;82(5):2020-2028.
Electrode fouling decreases sensitivity and can be a substantial limitation in electrochemical experiments. In this work we describe an electrochemical procedure that constantly renews the surface of a carbon microelectrode using periodic triangle voltage excursions to an extended anodic potential at a scan rate of 400 Vs−1. This methodology allows for the regeneration of an electrochemically active surface and restores electrode sensitivity degraded by irreversible adsorption of chemical species. We show that repeated voltammetric sweeps to moderate potentials in aqueous solution causes oxidative etching of carbon thereby constantly renewing the electrochemically active surface. Oxidative etching was established by tracking surface-localized fluorine atoms with XPS, by monitoring changes in carbon surface morphology with AFM on pyrolyzed photoresist films, and also by optical and electron microscopy. The use of waveforms with extended anodic potentials showed substantial increases in sensitivity towards the detection of catechols. This enhancement arose from the adsorption of the catechol moiety that could be maintained with a constant regeneration of the electrode surface. We also demonstrate that application of the extended waveform could restore the sensitivity of carbon microelectrodes diminished by irreversible adsorption (electrode fouling) of byproducts resulting from the electrooxidation and polymerization of tyramine. Overall, this work brings new insight into the factors that affect electrochemical processes at carbon electrodes and provides a simple method to remove or reduce fouling problems associated with many electrochemical experiments.
doi:10.1021/ac902753x
PMCID: PMC2838506  PMID: 20146453
electrode fouling; carbon-fiber microelectrode; pyrolyzed photoresist film; dopamine; catechols; adsorption
7.  Simultaneous Monitoring of Dopamine Concentration at Spatially Different Brain Locations in vivo 
Biosensors & bioelectronics  2009;25(5):1179.
When coupled with a microelectrode, background-subtracted fast scan cyclic voltammetry (FSCV) allows fast, sensitive and selective determination of analytes within a small spatial location. For the past 30 years experiments using this technique have been largely confined to recordings at a single microelectrode. Arrays with closely separated microelectrodes would allow researchers to gain more informative data as well as probe regions in close spatial proximity. This work presents one of the first FSCV microelectrode arrays (MEA) implemented in vivo with the ability to sample from different regions in close spatial proximity (equidistant within 1 mm). The array is manufactured from fused silica capillaries and a microfabricated electrode spacer. The functionality of the array is assessed by simultaneously monitoring electrically stimulated dopamine (DA) release in the striatum of anesthetized rat. As expected, heterogeneous dopamine release was simultaneously observed. Additionally, the pharmacological effect of raclopride (D2 receptor antagonist) and cocaine (monoamine uptake blocker) on the heterogeneity of DA release, in spatially different brain regions was shown to alter neurotransmitter release at all four electrode sites.
doi:10.1016/j.bios.2009.10.008
PMCID: PMC2818289  PMID: 19896822
8.  Simultaneous Decoupled Detection of Dopamine and Oxygen using Pyrolyzed Carbon Microarrays and FSCV 
Analytical chemistry  2009;81(15):6258-6265.
Microfabricated structures utilizing pyrolyzed photoresist have been shown to be a useful for monitoring electrochemical processes. These previous studies, however, were limited to constant potential measurements and slow scan voltammetry. Work described in this report utilizes microfabrication processes to produce devices that enable multiple fast-scan cyclic voltammetry (FSCV) waveforms to be applied to different electrodes on a single substrate. This enabled the simultaneous, decoupled, detection of dopamine and oxygen. This paper describes the fabrication process of these arrays and shows that pyrolyzed photoresist electrodes possess comparable surface chemistry and electrochemical properties to PAN type, T-650, carbon fiber microelectrodes using background-subtracted FSCV. The functionality of the array is discussed in terms of the degree of crosstalk in response to flow injections of physiologically relevant concentrations of dopamine and oxygen. Finally, other applications of pyrolyzed photoresist microelectrode arrays are shown, including: spatially resolved detection of analytes and combining FSCV with amperometry for the detection of dopamine.
doi:10.1021/ac900790m
PMCID: PMC2846216  PMID: 19552423
Pyrolyzed Photoresist; Microelectrode Arrays; Fast Scan Cyclic Voltammetry; Dopamine; Oxygen
9.  Electrochemical Dopamine Detection: Comparing Gold and Carbon Fiber Microelectrodes using Background Subtracted Fast Scan Cyclic Voltammetry 
Electrochemical detection is becoming increasingly important for the detection of biological species. Most current biological research with electrochemical detection is done with carbon fiber electrodes due to their many beneficial properties. The ability to build electrochemical sensor from noble metals instead of carbon fibers may be beneficial in developing inexpensive multiplexed electrochemical detection schemes. To advance understanding and to test the feasibility of using noble metal electrochemical sensors the detection of dopamine, a biologically important small molecule was studied here. Specifically, dopamine detection on gold microelectrodes was characterized and compared to P-55 carbon fiber microelectrodes of the same geometry, using background subtracted fast scan cyclic voltammetry. While not as sensitive to dopamine as carbon fibers, it was observed that gold microelectrodes have six times the saturation coverage per area and 40 times the linear working range. Selectivity to dopamine, in comparison to several other neurotransmitters and their derivatives, is also quantitatively described.
doi:10.1016/j.jelechem.2007.11.007
PMCID: PMC2659675  PMID: 19319208
10.  Activity of glucose oxidase functionalized onto magnetic nanoparticles 
Background
Magnetic nanoparticles have been significantly used for coupling with biomolecules, due to their unique properties.
Methods
Magnetic nanoparticles were synthesized by thermal co-precipitation of ferric and ferrous chloride using two different base solutions. Glucose oxidase was bound to the particles by direct attachment via carbodiimide activation or by thiophene acetylation of magnetic nanoparticles. Transmission electron microscopy was used to characterize the size and structure of the particles while the binding of glucose oxidase to the particles was confirmed using Fourier transform infrared spectroscopy.
Results
The direct binding of glucose oxidase via carbodiimide activity was found to be more effective, resulting in bound enzyme efficiencies between 94–100% while thiophene acetylation was 66–72% efficient. Kinetic and stability studies showed that the enzyme activity was more preserved upon binding onto the nanoparticles when subjected to thermal and various pH conditions. The overall activity of glucose oxidase was improved when bound to magnetic nanoparticles
Conclusion
Binding of enzyme onto magnetic nanoparticles via carbodiimide activation is a very efficient method for developing bioconjugates for biological applications
doi:10.1186/1477-044X-3-1
PMCID: PMC555562  PMID: 15762994
11.  Examination of Cholesterol oxidase attachment to magnetic nanoparticles 
Magnetic nanoparticles (Fe3O4) were synthesized by thermal co-precipitation of ferric and ferrous chlorides. The sizes and structure of the particles were characterized using transmission electron microscopy (TEM). The size of the particles was in the range between 9.7 and 56.4 nm. Cholesterol oxidase (CHO) was successfully bound to the particles via carbodiimide activation. FTIR spectroscopy was used to confirm the binding of CHO to the particles. The binding efficiency was between 98 and 100% irrespective of the amount of particles used. Kinetic studies of the free and bound CHO revealed that the stability and activity of the enzyme were significantly improved upon binding to the nanoparticles. Furthermore, the bound enzyme exhibited a better tolerance to pH, temperature and substrate concentration. The activation energy for free and bound CHO was 13.6 and 9.3 kJ/mol, respectively. This indicated that the energy barrier of CHO activity was reduced upon binding onto Fe3O4 nanoparticles. The improvements observed in activity, stability, and functionality of CHO resulted from structural and conformational changes of the bound enzyme. The study indicates that the stability and activity of CHO could be enhanced via attachment to magnetic nanoparticles and subsequently will contribute to better uses of this enzyme in various biological and clinical applications.
doi:10.1186/1477-3155-3-1
PMCID: PMC548673  PMID: 15661076

Results 1-11 (11)