Thesis advisor: Michael J. Naughton / Thesis advisor: Michael J. Burns / Recent progress in the study of the brain has been greatly facilitated by the development of new measurement tools capable of minimally-invasive, robust coupling to neuronal assemblies. Two prominent examples are the microelectrode array, which enables electrical signals from large numbers of neurons to be detected and spatiotemporally correlated, and optogenetics, which enables the electrical activity of cells to be controlled with light. In the former case, high spatial density is desirable but, as electrode arrays evolve toward higher density and thus smaller pitch, electrical crosstalk increases. In the latter, finer control over light input is desirable, to enable improved studies of neuroelectronic pathways emanating from specific cell stimulation. Herein, we introduce a coaxial electrode architecture that is uniquely suited to address these issues, as it can simultaneously be utilized as an optical waveguide and a shielded electrode in dense arrays. / Thesis (PhD) — Boston College, 2017. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Physics.

To understand how the brain functions and what disruptions underlie neurological diseases and disorders, analytical methods are needed that can succeed in the complexity of the native brain environment. To make a measurement in functioning, live tissue, these methods must be selective for specific analytes in a matrix that has over 1000 different chemical species, be able to measure chemical changes on multiple timescales (10-3 s to 104 s), have a high spatial resolution (μm), and be sensitive (pM to μM). The work described within, details the development and application of a voltammetric method, fast-scan controlled adsorption voltammetry (FSCAV) that is capable of monitoring baseline levels of serotonin and dopamine, as well as monitoring changes on multiple time scales with high sensitivity and selectivity. Because FSCAV is performed using a carbon-fiber microelectrode, the same sensor can be used for fast-scan cyclic voltammetry to monitor rapid (phasic) changes of dopamine and serotonin in the extracellular space. Thus a single-sensor strategy for measuring tonic and phasic concentrations of these important neurotransmitters is developed and used to elucidate important insight into the differences of serotonin and dopamine regulation. Additionally it is revealed that dopamine exhibits a coaction between tonic and phasic signaling where serotonin does not. Using this approach, a method for evaluating pain processing in a preclinical model is developed, which reveals an important relationship between chronic pain and dopamine signaling. Furthermore, a mathematical model to describe mass-transport limited adsorption is developed and used to determine the diffusion coefficient of both dopamine and serotonin in situ. The work described within details an important advancement in neuroanalytical methodology that will provide new insights both short-term and long-term for studying fundamental chemical mechanisms of neurotransmission.

Microelectrodes have been found to be a valuable tool in a variety of analytical studies. Their advantages over macro-sized electrodes are well known, including their enhanced mass transport properties (due to their ubiquitous hemispherical diffusion) which lead to steady state responses without external convection. They also exhibit high signal-to-noise ratios (greater sensitivities), furthering their analytical application. Microelectrode arrays are analytical devices with multiple electrodes. There are suitable for practical sensing with all the benefits of microelectrodes but with greater currents, leading to greater ease of measurement. To produce a reliable electroanalytical device the microelectrode response must be reproducible, a fundamental property based on the quality control of their production. Square microelectrode and array fabrication techniques have been developed for this purpose. This research discusses the fabrication and development of closely spaced arrays of square microelectrodes. Simulated and measured responses are compared and used to characterize electrode and array responses by cyclic voltammetry, electrical impedance spectroscopy and current-time transients. Measurements on variably spaced arrays allow insight into overlap of hemispherical diffusion from individual electrodes and the subsequent effect including peak current output on the array device. By studying these devices key insights into the mass transport properties of single square microelectrodes and microelectrode arrays were gained. This study also prepares and develops microelectrodes from materials appropriate for use in the extreme environments of molten salts and concentrated nitric acid solutions. These robust electrodes were developed for use in hydro- and pyro-chemical techniques for nuclear fuel reprocessing. These results demonstrate the practical uses for microelectrode systems across a wide range of chemical systems and in extreme conditions.

Thesis (Ph.D)--Biomedical Engineering, Georgia Institute of Technology, 2009. / Committee Chair: Bellamkonda, Ravi; Committee Member: Babensee, Julia; Committee Member: Butera, Robert; Committee Member: DeWeerth, Steve; Committee Member: Lee, Robert; Committee Member: McKeon, Robert. Part of the SMARTech Electronic Thesis and Dissertation Collection.

A report from the National Institutes of Health defines a disease biomarker as a "characteristic that is objectively measured and evaluated as an indicator of normal biologic processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention." Early diagnosis is a crucial factor for incurable disease such as cancer and Alzheimer’s disease (AD). During the last decade researchers have discovered that biochemical changes caused by a disease can be detected considerably earlier as compared to physical manifestations/symptoms. In this dissertation electrochemical detection was utilized as the detection strategy as it offers high sensitivity/specificity, ease of operation, and capability of miniaturization and multiplexed detection. Electrochemical detection of biological analytes is an established field, and has matured at a rapid pace during the last 50 years and adapted itself to advances in micro/nanofabrication procedures. Carbon fiber microelectrodes were utilized as the platform sensor due to their high signal to noise ratio, ease and low-cost of fabrication, biocompatibility, and active carbon surface which allows conjugation with bio-recognition moieties. This dissertation specifically focuses on the detection of 3 extensively validated biomarkers for cancer and AD. Firstly, vascular endothelial growth factor (VEGF) a cancer biomarker was detected using a one-step, reagentless immunosensing strategy. The immunosensing strategy allowed a rapid and sensitive means of VEGF detection with a detection limit of about 38 pg/mL with a linear dynamic range of 0–100 pg/mL. Direct detection of AD-related biomarker amyloid beta (Aβ) was achieved by exploiting its inherent electroactivity. The quantification of the ratio of Aβ1-40/42 (or Aβ ratio) has been established as a reliable test to diagnose AD through human clinical trials. Triple barrel carbon fiber microelectrodes were used to simultaneously detect Aβ1-40 and Aβ1-42 in cerebrospinal fluid from rats within a detection range of 100nM to 1.2μM and 400nM to 1μM respectively. In addition, the release of DNA damage/repair biomarker 8-hydroxydeoxyguanine (8-OHdG) under the influence of reactive oxidative stress from single lung endothelial cell was monitored using an activated carbon fiber microelectrode. The sensor was used to test the influence of nicotine, which is one of the most biologically active chemicals present in cigarette smoke and smokeless tobacco.

No description available.

No description available.

The prefrontal cortex (PFC) is an area of the brain that is critically important for learning, memory, organization, and integration, and PFC dysfunction has been associated with pathologies including Alzheimer’s disease, schizophrenia, and drug addiction. However, there exists a paucity of information regarding neurochemical signaling in the distinct sub-regions of the PFC, particularly the medial prefrontal cortex (mPFC). The mPFC receives glutamatergic input from a number of brain areas, and functional glutamate signaling is essential for normal cognitive processes. To further understand glutamate neurotransmission, in vivo measurements of glutamate were performed in the cingulate cortex, prelimbic cortex, and infralimbic cortex of anesthetized rats using enzyme-based microelectrode array technology. Measurements of acetylcholine were also performed to examine the relationship between glutamate and other neurotransmitters in the mPFC. The described studies revealed a homogeneity of glutamate and acetylcholine signaling in the mPFC sub-regions, indicating somewhat uniform tonic and phasic levels of these two transmitters. In the infralimbic mPFC of awake freely-moving rats, rapid, phasic glutamate signaling events, termed “transients” were observed and in vivo glutamate signaling was successfully monitored over 24 hour time periods. The effects of methylphenidate (MPH), a stimulant medication with abuse potential that is used in the treatment of attention-deficit hyperactivity disorder, were measured in mPFC sub-regions of anesthetized rats. Data revealed similar tonic and phasic glutamate levels between chronic MPH-treated rats and controls in all sub-regions. Locomotor data from the chronic treatment period supported the behavioral sensitization effects of multiple MPH treatments. Significant effects were observed in locomotor activity, resting levels of glutamate, and glutamate uptake rates in the infralimbic mPFC of awake, freely-moving animals that received chronic MPH treatment. Taken together, this body of work characterizes glutamate signaling in the rat mPFC to a degree never before reported, and serves to report for the first time the effects of MPH on glutamate signaling in the mPFC.

No presente trabalho, foi utilizada a microscopia eletroquímica de varredura (SECM) com o intuito de obter informações sobre a topografia, reatividade e possíveis intermediários formados em superfícies eletródicas. Foram estudados, em uma etapa inicial, modelos que proporcionaram a aprendizagem do manuseio e tratamento de dados utilizando a SECM. Em uma segunda etapa, a SECM foi aplicada no estudo da reatividade de substratos de cobre que passaram por um processo de ativação eletroquímica. A ativação foi realizada imergindo uma superfície de cobre em uma solução de Na2SO4 0,1 mol L-1 (pH = 2) e posterior aplicação de pulsos de potencial para oxidação de cobre (500 mV vs. Ag/AgCl(KCl sat.)) e subseqüente redução de íons Cu(II) (-250 mV vs. Ag/AgCl(KCl sat.)). Esse procedimento confere um aumento na sensibilidade na determinação de nitrato e nitrito. Para o estudo foram obtidas curvas de aproximação as quais foram correlacionadas com o tempo empregado na ativação. Também foram obtidas informações sobre a geração de Cu(I), estabilizado na presença de íons cloreto, durante o processo de ativação e sua influência na determinação de nitrato e nitrito. Por fim, a SECM foi utilizada no estudo de um sistema biomimético de cobre que catalisa a redução de oxigênio. Foram obtidas imagens com informações sobre a reatividade da superfície, homogeneidade do recobrimento do substrato e o mecanismo da reação. / Scanning electrochemical microscopy (SECM) was employed in order to obtain information on the topography, reactivity, and possible intermediates generated at electrodic surfaces. Standard chemical systems were firstly investigated as an attempt to understand the principles of the technique. Then, SECM was used to study the reactivity of copper substrates upon activation by an electrochemical protocol. The surface activation was achieved by immersing a copper surface in a solution of 0.1 mol L-1 Na2SO4 (pH = 2) for further anodic oxidation of copper (500 mV vs. Ag / AgCl(sat. KCl)) and subsequent reduction of Cu (II) (-250 mV vs. Ag / AgCl(sat. KCl)). An enhanced sensitivity was noticed for the determination of nitrate and nitrite at the modified copper surface and the approach curves were correlated with the activation time. Information on the generation of Cu (I), which was stabilized in the presence of chloride ions during the activation process, was also obtained, as well as the influence of this intermediate on the determination of nitrate and nitrite. Finally, SECM was used in the study of a copper biomimetic system which is able to catalyze the reduction of oxygen. Information on the reactivity of the surface, the homogeneity of the coated substrate and the reaction mechanism were obtained by analyzing the SECM results.

No presente trabalho, foi utilizada a microscopia eletroquímica de varredura (SECM) com o intuito de obter informações sobre a topografia, reatividade e possíveis intermediários formados em superfícies eletródicas. Foram estudados, em uma etapa inicial, modelos que proporcionaram a aprendizagem do manuseio e tratamento de dados utilizando a SECM. Em uma segunda etapa, a SECM foi aplicada no estudo da reatividade de substratos de cobre que passaram por um processo de ativação eletroquímica. A ativação foi realizada imergindo uma superfície de cobre em uma solução de Na2SO4 0,1 mol L-1 (pH = 2) e posterior aplicação de pulsos de potencial para oxidação de cobre (500 mV vs. Ag/AgCl(KCl sat.)) e subseqüente redução de íons Cu(II) (-250 mV vs. Ag/AgCl(KCl sat.)). Esse procedimento confere um aumento na sensibilidade na determinação de nitrato e nitrito. Para o estudo foram obtidas curvas de aproximação as quais foram correlacionadas com o tempo empregado na ativação. Também foram obtidas informações sobre a geração de Cu(I), estabilizado na presença de íons cloreto, durante o processo de ativação e sua influência na determinação de nitrato e nitrito. Por fim, a SECM foi utilizada no estudo de um sistema biomimético de cobre que catalisa a redução de oxigênio. Foram obtidas imagens com informações sobre a reatividade da superfície, homogeneidade do recobrimento do substrato e o mecanismo da reação. / Scanning electrochemical microscopy (SECM) was employed in order to obtain information on the topography, reactivity, and possible intermediates generated at electrodic surfaces. Standard chemical systems were firstly investigated as an attempt to understand the principles of the technique. Then, SECM was used to study the reactivity of copper substrates upon activation by an electrochemical protocol. The surface activation was achieved by immersing a copper surface in a solution of 0.1 mol L-1 Na2SO4 (pH = 2) for further anodic oxidation of copper (500 mV vs. Ag / AgCl(sat. KCl)) and subsequent reduction of Cu (II) (-250 mV vs. Ag / AgCl(sat. KCl)). An enhanced sensitivity was noticed for the determination of nitrate and nitrite at the modified copper surface and the approach curves were correlated with the activation time. Information on the generation of Cu (I), which was stabilized in the presence of chloride ions during the activation process, was also obtained, as well as the influence of this intermediate on the determination of nitrate and nitrite. Finally, SECM was used in the study of a copper biomimetic system which is able to catalyze the reduction of oxygen. Information on the reactivity of the surface, the homogeneity of the coated substrate and the reaction mechanism were obtained by analyzing the SECM results.