In silico study on in vitro experiments to determine the electric membrane properties of a realistic cochlear model for electric field simulations on cochlear implants

To further develop and optimise the design of cochlear implants, a numerical model with precise material properties and authentic geometry is required. Since simulation results strongly depend on the accuracy of the estimates of the electrical properties of cochlear membranes, it is important to have a reliable in vivo method for measuring electrical impedance changes in the cochlear compartments. This work is a preliminary attempt to model, simulate and analyse the behaviour of a novel in-vitro experimental system for conducting plausible in-vivo measurements on mammalian cochlea membranes.Zur Weiterentwicklung und Optimierung des Designs von Cochlea-Implantaten ist ein detailliertes numerisches Modell der Cochlea erforderlich. Da die Simulationsergebnisse stark von den elektrischen Eigenschaften der Cochlea-Membranen abhängen, ist es wichtig, ein zuverlässiges In-vivo-Verfahren zur Messung des elektrischen Impedanzverlaufs zu haben. Diese Arbeit ist eine vorbereitende Studie, das Verhalten eines neuartigen In-vitro-Versuchssystems zur Durchführung plausibler In-vivo-Messungen an Cochlea-Membranen von Säugetieren zu modellieren, zu simulieren und zu analysieren

Functionalized graphene sensors for real time monitoring fermentation processes:electrochemical and chemiresistive sensors

We developed a reference-less, chemiresistive, solid-state pH sensor to determine the acidification of the fermentation liquid in real time during the growth of Lactococcus lactis. One of the crucial findings of this work was that the ERGO-PA could not be used as such. It appeared that it was necessary to protect the sensor area with a Nafion coating to measure the pH in the fermentation broth. Most likely, the change in the concentration of redox-active components in the fermentation broth influences the conductivity of the ERGO-PA. Nafion formed a cation-selective membrane on top of the ERGO-PA allowing protons to diffuse to the selective layer of the sensor but not the redox-active components in the fermentation medium. We also reported a new approach to measure the dissolved oxygen concentration (DO) in a fermentation broth. The functionality of the sensor to measure DO was demonstrated during the growth of the obligate aerobic actinomycete Amycalotopsis methanolica in miniaturized 3D-printed bioreactors. For this oxygen-sensing application, the required modifications were obtained by doping hydrothermally reduced graphene oxide with nitrogen and boron atoms (N,B-HRGO). Further, these chemiresistive sensors are housed in the 3D printed bioreactor lid and used to measure pH, DO, and biomass in 3 ml fermentation broth. Additionally, the pH-sensor was equipped with a small heating element and a temperature sensor and that could be used for temperature control of the fermentation liquid. The setup was demonstrated to measure the pH, DO, temperature and biomass concentration in four parallel bioreactors

Development and modelling of a versatile active micro-electrode array for high density in-vivo and in-vitro neural signal investigation

The electrophysiological observation of neurological cells has allowed much knowledge to be gathered regarding how living organisms are believed to acquire and process sensation. Although much has been learned about neurons in isolation, there is much more to be discovered in how these neurons communicate within large networks. The challenges of measuring neurological networks at the scale, density and chronic level of non invasiveness required to observe neurological processing and decision making are manifold, however methods have been suggested that have allowed small scale networks to be observed using arrays of micro-fabricated electrodes. These arrays transduce ionic perturbations local to the cell membrane in the extracellular fluid into small electrical signals within the metal that may be measured. A device was designed for optimal electrical matching to the electrode interface and maximal signal preservation of the received extracellular neural signals. Design parameters were developed from electrophysiological computer simulations and experimentally obtained empirical models of the electrode-electrolyte interface. From this information, a novel interface based signal filtering method was developed that enabled high density amplifier interface circuitry to be realised. A novel prototype monolithic active electrode was developed using CMOS microfabrication technology. The device uses the top metallization of a selected process to form the electrode substrate and compact amplification circuitry fabricated directly beneath the electrode to amplify and separate the neural signal from the baseline offsets and noise of the electrode interface. The signal is then buffered for high speed sampling and switched signal routing. Prototype 16 and 256 active electrode array with custom support circuitry is presented at the layout stage for a 20 μm diameter 100 μm pitch electrode array. Each device consumes 26.4 μW of power and contributes 4.509 μV (rms) of noise to the received signal over a controlled bandwidth of 10 Hz - 5 kHz. The research has provided a fundamental insight into the challenges of high density neural network observation, both in the passive and the active manner. The thesis concludes that power consumption is the fundamental limiting factor of high density integrated MEA circuitry; low power dissipation being crucial for the existence of the surface adhered cells under measurement. With transistor sizing, noise and signal slewing each being inversely proportional to the dc supply current and the large power requirements of desirable ancillary circuitry such as analogue-to-digital converters, a situation of compromise is approached that must be carefully considered for specific application design

Brain-Computer Interfaces using Electrocorticography and Surface Stimulation

The brain connects to, modulates, and receives information from every organ in the body. As such, brain-computer interfaces (BCIs) have vast potential for diagnostics, medical therapies, and even augmentation or enhancement of normal functions. BCIs provide a means to explore the furthest corners of what it means to think, to feel, and to act—to experience the world and to be who you are. This work focuses on the development of a chronic bi-directional BCI for sensorimotor restoration through the use of separable frequency bands for recording motor intent and providing sensory feedback via electrocortical stimulation. Epidural cortical surface electrodes are used to both record electrocorticographic (ECoG) signals and provide stimulation without adverse effects associated with penetration through the protective dural barrier of brain. Chronic changes in electrode properties and signal characteristics are discussed, which inform optimal electrode designs and co-adaptive algorithms for decoding high-dimensional information. Additionally, a multi-layered approach to artifact suppression is presented, which includes a systems-level design of electronics, signal processing, and stimulus waveforms. The results of this work are relevant to a wider range of applications beyond ECoG and BCIs that involve closed-loop recording and stimulation throughout the body. By enabling simultaneous recording and stimulation through the techniques described here, responsive therapies can be developed that are tuned to individual patients and provide precision therapies at exactly the right place and time. This has the potential to improve targeted therapeutic outcomes while reducing undesirable side effects

Electrochemical biosensor based on microfabricated electrode arrays for life sciences applications

In developing a biosensor, the utmost important aspects that need to be emphasized are the specificity and selectivity of the transducer. These two vital prerequisites are of paramount in ensuring a robust and reliable biosensor. Improvements in electrochemical sensors can be achieved by using microelectrodes and to modify the electrode surface (using chemical or biological recognition layers to improve the sensitivity and selectivity). The fabrication and characterisations of silicon-based and glass-based gold microelectrode arrays with various geometries (band and disc) and dimension (ranging from 10 μm-100 nm) were reported. It was found that silicon-based transducers of 10 μm gold microelectrode array exhibited the most stable and reproducible electrochemical measurements hence this dimension was selected for further study. Chemical electrodeposition on both 10 μm microband and microdisc were found viable by electro-assisted self-assembled sol-gel silica film and nanoporous-gold electrodeposition respectively. The fabrication and characterisations of on-chip electrochemical cell was also reported with a fixed diameter/width dimension and interspacing variation. With this regard, the 10 μm microelectrode array with interspacing distance of 100 μm exhibited the best electrochemical response. Surface functionalisations on single chip of planar gold macroelectrodes were also studied for the immobilisation of histidine-tagged protein and antibody. Imaging techniques such as atomic force microscopy, fluorescent microscopy or scanning electron microscope were employed to complement the electrochemical characterisations. The long-chain thiol of self-assembled monolayer with NTA-metal ligand coordination was selected for the histidine-tagged protein while silanisation technique was selected for the antibody immobilisation. The final part of the thesis described the development of a T-2 labelless immunosensor using impedimetric approach. Good antibody calibration curve was obtained for both 10 μm microband and 10 μm microdisc array. For the establishment of the T-2/HT-2 toxin calibration curve, it was found that larger microdisc array dimension was required to produce better calibration curve. The calibration curves established in buffer solution show that the microelectrode arrays were sensitive and able to detect levels of T-2/HT-2 toxin as low as 25 ppb (25 μg kg-1) with a limit of quantitation of 4.89 ppb for a 10 μm microband array and 1.53 ppb for the 40 μm microdisc array

Optimal Electrode Size for Multi-Scale Extracellular-Potential Recording From Neuronal Assemblies

Advances in microfabrication technology have enabled the production of devices containing arrays of thousands of closely spaced recording electrodes, which afford subcellular resolution of electrical signals in neurons and neuronal networks. Rationalizing the electrode size and configuration in such arrays demands consideration of application-specific requirements and inherent features of the electrodes. Tradeoffs among size, spatial density, sensitivity, noise, attenuation, and other factors are inevitable. Although recording extracellular signals from neurons with planar metal electrodes is fairly well established, the effects of the electrode characteristics on the quality and utility of recorded signals, especially for small, densely packed electrodes, have yet to be fully characterized. Here, we present a combined experimental and computational approach to elucidating how electrode size, and size-dependent parameters, such as impedance, baseline noise, and transmission characteristics, influence recorded neuronal signals. Using arrays containing platinum electrodes of different sizes, we experimentally evaluated the electrode performance in the recording of local field potentials (LFPs) and extracellular action potentials (EAPs) from the following cell preparations: acute brain slices, dissociated cell cultures, and organotypic slice cultures. Moreover, we simulated the potential spatial decay of point-current sources to investigate signal averaging using known signal sources. We demonstrated that the noise and signal attenuation depend more on the electrode impedance than on electrode size, per se, especially for electrodes <10 μm in width or diameter to achieve high-spatial-resolution readout. By minimizing electrode impedance of small electrodes (<10 μm) via surface modification, we could maximize the signal-to-noise ratio to electrically visualize the propagation of axonal EAPs and to isolate single-unit spikes. Due to the large amplitude of LFP signals, recording quality was high and nearly independent of electrode size. These findings should be of value in configuring in vitro and in vivo microelectrode arrays for extracellular recordings with high spatial resolution in various applications

EXPLORING THE ROLE AND IMPACT OF MICROSCALE PHENOMENA ON ELECTRODE, MICRODEVICE, AND CELLULAR FUNCTION

Microfluidic technologies enable the development of portable devices to perform multiple high-resolution unit operations with small sample and reagent volumes, low fabrication cost, facile operation, and quick response times. Microfluidic platforms are expected to effectively interpret both wanted and unwanted phenomena; however, a comprehensive evaluation of the unwanted phenomena has not been appropriately investigated in the literature. This work explored an attenuative evaluation of unwanted phenomena, also called here as secondary phenomena, in a unique approach. Upon electric field utilization within microfluidic devices, electrode miniaturization improves device sensitivity. However, electrodes in contact with medium solution can yield byproducts that can change medium properties such as pH as well as bulk ion concentration and eventually target cell viability. While electrode byproducts are sometimes beneficial; but, this is not always the case. Two strategies were employed to protect cells from the electrode byproducts: (i) coating the electrodes with hafnium oxide (HfO2), and (ii) stabilization of the cell membrane using a low concentration of Triton X-100 surfactant. Our results showed that both strategies are a plausible way to selectively isolate cell and reduce the risk of contamination from electrode byproducts. The design of a medium solution is also critical to minimize unwanted cell-medium interaction. Surfactants are frequently added to cell-medium solutions to improve sensitivity and reproducibility without disrupting protein composition of cell membranes or cell viability. In non-electrokinetic systems, surfactants have been shown to reduce interfacial tensions and prevent analyte sticking. However, the impacts of surfactant interactions with cell membranes have not previously been explored in electrokinetic systems. This work indicated the dynamic surfactant interactions with cell membranes which altered the cell membrane integrity. It is important that the effects of the chemical interactions between cells to be fully explored and to be separately attributed to reported cellular responses to accurate catalog properties and engineer reliable microfluidic electrokinetic devices. Finally, a comprehensive level of understanding led us to utilize dielectrophoresis in its full capacity as a tool to monitor the state and progression of virus infection as well as anti-viral activities of regenerative compound. Glycine was utilized as potential antiviral compounds to reduce porcine parvovirus (PPV) infection in porcine kidney (PK-13) cells. Our results demonstrate that the glycine altered the virus-host interactions during virus assembly. Thus, elucidating the mechanisms of these novel antiviral compounds is crucial to their development as potential therapeutic drugs