92F05 Other natural sciences (should also be assigned at least one other classification number in section 92)
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In the present thesis, AC electrokinetic forces, like dielectrophoresis and AC electroosmosis, were demonstrated as a simple and fast method to functionalize the surface of nanoelectrodes with submicrometer sized biological objects. These nanoelectrodes have a cylindrical shape with a diameter of 500 nm arranged in an array of 6256 electrodes. Due to its medical relevance influenza virus as well as anti-influenza antibodies were chosen as a model organism. Common methods to bring antibodies or proteins to biosensor surfaces are complex and time-consuming. In the present work, it was demonstrated that by applying AC electric fields influenza viruses and antibodies can be immobilized onto the nanoelectrodes within seconds without any prior chemical modification of neither the surface nor the immobilized biological object. The distribution of these immobilized objects is not uniform over the entire array, it exhibits a decreasing gradient from the outer row to the inner ones. Different causes for this gradient have been discussed, such as the vortex-shaped fluid motion above the nanoelectrodes generated by, among others, electrothermal fluid flow. It was demonstrated that parts of the accumulated material are permanently immobilized to the electrodes. This is a unique characteristic of the presented system since in the literature the AC electrokinetic immobilization is almost entirely presented as a method just for temporary immobilization. The spatial distribution of the immobilized viral material or the anti-influenza antibodies at the electrodes was observed by either the combination of fluorescence microscopy and deconvolution or by super-resolution microscopy (STED). On-chip immunoassays were performed to examine the suitability of the functionalized electrodes as a potential affinity-based biosensor. Two approaches were pursued: A) the influenza virus as the bio-receptor or B) the influenza virus as the analyte. Different sources of error were eliminated by ELISA and passivation experiments. Hence, the activity of the immobilized object was inspected by incubation with the analyte. This resulted in the successful detection of anti-influenza antibodies by the immobilized viral material. On the other hand, a detection of influenza virus particles by the immobilized anti-influenza antibodies was not possible. The latter might be due to lost activity or wrong orientation of the antibodies. Thus, further examinations on the activity of by AC electric fields immobilized antibodies should follow. When combined with microfluidics and an electrical read-out system, the functionalized chips possess the potential to serve as a rapid, portable, and cost-effective point-of-care (POC) device. This device can be utilized as a basis for diverse applications in diagnosing and treating influenza, as well as various other pathogens.
This work develops hybrid methods of imaging spectroscopy for open pit mining and examines their feasibility compared with state-of-the-art. The material distribution within a mine face differs in the small scale and within daily assigned extraction segments. These changes can be relevant to subsequent processing steps but are not always visually identifiable prior to the extraction. Misclassifications that cause false allocations of extracted material need to be minimized in order to reduce energy-intensive material re-handling. The use of imaging spectroscopy aspires to the allocation of relevant deposit-specific materials before extraction, and allows for efficient material handling after extraction. The aim of this work is the parameterization of imaging spectroscopy for pit mining applications and the development and evaluation of a workflow for a mine face, ground- based, spectral characterization. In this work, an application-based sensor adaptation is proposed. The sensor complexity is reduced by down-sampling the spectral resolution of the system based on the samples’ spectral characteristics. This was achieved by the evaluation of existing hyperspectral outcrop analysis approaches based on laboratory sample scans from the iron quadrangle in Minas Gerais, Brazil and by the development of a spectral mine face monitoring workflow which was tested for both an operating and an inactive open pit copper mine in the Republic of Cyprus.
The workflow presented here is applied to three regional data sets: 1) Iron ore samples from Brazil, (laboratory); 2) Samples and hyperspectral mine face imagery from the copper-gold-pyrite mine Apliki, Republic of Cyprus (laboratory and mine face data); and 3) Samples and hyperspectral mine face imagery from the copper-gold-pyrite deposit Three Hills, Republic of Cyprus (laboratory and mine face data). The hyperspectral laboratory dataset of fifteen Brazilian iron ore samples was used to evaluate different analysis methods and different sensor models. Nineteen commonly used methods to analyze and map hyperspectral data were compared regarding the methods’ resulting data products and the accuracy of the mapping and the analysis computation time. Four of the evaluated methods were determined for subsequent analyses to determine the best-performing algorithms: The spectral angle mapper (SAM), a support vector machine algorithm (SVM), the binary feature fitting algorithm (BFF) and the EnMap geological mapper (EnGeoMap). Next, commercially available imaging spectroscopy sensors were evaluated for their usability in open pit mining conditions. Step-wise downsampling of the data - the reduction of the number of bands with an increase of each band’s bandwidth - was performed to investigate the possible simplification and ruggedization of a sensor without a quality fall-off of the mapping results. The impact of the atmosphere visible in the spectrum between 1300–2010nm was reduced by excluding the spectral range from the data for mapping. This tested the feasibility of the method under realistic open pit data conditions. Thirteen datasets based on the different, downsampled sensors were analyzed with the four predetermined methods. The optimum sensor for spectral mine face material distinction was determined as a VNIR-SWIR sensor with 40nm bandwidths in the VNIR and 15nm bandwidths in the SWIR spectral range and excluding the atmospherically impacted bands. The Apliki mine sample dataset was used for the application of the found optimal analyses and sensors. Thirty-six samples were analyzed geochemically and mineralogically. The sample spectra were compiled to two spectral libraries, both distinguishing between seven different geochemical-spectral clusters. The reflectance dataset was downsampled to five different sensors. The five different datasets were mapped with the SAM, BFF and SVM method achieving mapping accuracies of 85-72%, 85-76% and 57-46% respectively. One mine face scan of Apliki was used for the application of the developed workflow. The mapping results were validated against the geochemistry and mineralogy of thirty-six documented field sampling points and a zonation map of the mine face which is based on sixty-six samples and field mapping. The mine face was analyzed with SAM and BFF. The analysis maps were visualized on top of a Structure-from-Motion derived 3D model of the open pit. The mapped geological units and zones correlate well with the expected zonation of the mine face. The third set of hyperspectral imagery from Three Hills was available for applying the fully-developed workflow. Geochemical sample analyses and laboratory spectral data of fifteen different samples from the Three Hills mine, Republic of Cyprus, were used to analyse a downsampled mine face scan of the open pit. Here, areas of low, medium and high ore content were identified.
The developed workflow is successfully applied to the open pit mines Apliki and Three Hills and the spectral maps reflect the prevailing geological conditions. This work leads through the acquisition, preparation and processing of imaging spectroscopy data, the optimum choice of analysis methodology, and the utilization of simplified, robust sensors that meet the requirements of open pit mining conditions. It accentuates the importance of a site-specific and deposit-specific spectral library for the mine face analysis and underlines the need for geological and spectral analysis experts to successfully implement imaging spectroscopy in the field of open pit mining.
Electron transfer (ET) reactions play a crucial role in the metabolic pathways of all organisms. In biotechnological approaches, the redox properties of the protein cytochrome c (cyt c), which acts as an electron shuttle in the respiratory chain, was utilized to engineer ET chains on electrode surfaces. With the help of the biopolymer DNA, the redox protein assembles into electro active multilayer (ML) systems, providing a biocompatible matrix for the entrapment of proteins.
In this study the characteristics of the cyt c and DNA interaction were defined on the molecular level for the first time and the binding sites of DNA on cyt c were identified. Persistent cyt c/DNA complexes were formed in solution under the assembly conditions of ML architectures, i.e. pH 5.0 and low ionic strength. At pH 7.0, no agglomerates were formed, permitting the characterization of the NMR spectroscopy. Using transverse relaxation-optimized spectroscopy (TROSY)-heteronuclear single quantum coherence (HSQC) experiments, DNAs’ binding sites on the protein were identified. In particular, negatively charged AA residues, which are known interaction sites in cyt c/protein binding were identified as the main contact points of cyt c and DNA.
Moreover, the sophisticated task of arranging proteins on electrode surfaces to create functional ET chains was addressed. Therefore, two different enzyme types, the flavin dependent fructose dehydrogenase (FDH) and the pyrroloquinoline quinone dependent glucose dehydrogenase (PQQ-GDH), were tested as reaction partners of freely diffusing cyt c and cyt c immobilized on electrodes in mono- and MLs. The characterisation of the ET processes was performed by means of electrochemistry and the protein deposition was monitored by microgravimetric measurements. FDH and PQQ-GDH were found to be generally suitable for combination with the cyt c/DNA ML system, since both enzymes interact with cyt c in solution and in the immobilized state. The immobilization of FDH and cyt c was achieved with the enzyme on top of a cyt c monolayer electrode without the help of a polyelectrolyte. Combining FDH with the cyt c/DNA ML system did not succeed, yet. However, the basic conditions for this protein-protein interaction were defined. PQQ-GDH was successfully coupled with the ML system, demonstrating that that the cyt c/DNA ML system provides a suitable interface for enzymes and that the creation of signal chains, based on the idea of co-immobilized proteins is feasible.
Future work may be directed to the investigation of cyt c/DNA interaction under the precise conditions of ML assembly. Therefore, solid state NMR or X-ray crystallography may be required. Based on the results of this study, the combination of FDH with the ML system should be addressed. Moreover, alternative types of enzymes may be tested as catalytic component of the ML assembly, aiming on the development of innovative biosensor applications.