|
|
Staphylococcus aureus - jeho výskyt a možnosti detekce v mléce a mléčných výrobcích
Fenclová, Denisa
The theoretical part of diploma thesis is focused on summarizing current knowledge about S. aureus, its pathogenesis, occurrence, and possibilities of its detection in milk and dairy products. The experimental part of diploma thesis is aimed at determining S. aureus using various cultivation methods. S. aureus was determined using B-P agar with the addition of RPF, ClearMilku test, MicroFast®, and PetrifilmTM. The determination was evaluated as follows: B-P agar with the addition of RPF > PetrifilmTM > MicroFast® > ClearMilk test. Another part of the experimental diploma thesis deals with the possibility of detecting S. aureus using a newly developed biosensor detection system.
|
|
| |
|
|
Nanopatterned alumina-based materials for electrochemical sensors and biosensors
Kynclová, Hana ; Hynek, David (referee) ; Trnková, Libuše (referee) ; Prášek, Jan (advisor)
The doctoral thesis is focused on basic research and development of nanostructured surfaces prepared using anodic alumina material. Various types of gold nanostructured surfaces and nanoporous aluminum membranes for electrochemical sensors and biosensors were prepared using the anodic oxidation method. Nanostructured surfaces were prepared by electrochemical anodization of aluminum material to form hexagonally arranged nanopores. Gold was then deposited into the nanoporous masks by electrochemical reduction from potassium dicyanoaurate solution using a pulse deposition method. The prepared nanostructured gold surfaces were electrochemically characterized by electrochemical impedance spectroscopy and voltammetry. Temperature stability and the effect of annealing on their electrochemical behavior at atmospheric pressure as well as in the vacuum were investigated. Then, gold nanostructures of various dimensions were prepared and the influence of their shape and dimensions on the electrochemical behavior was studied. Nanostructured surfaces were also modified with 11–mercaptoundecanoic acid, and the effect of this modification on the electrochemical results was studied. In the last part of the work, nanoporous aluminum membranes were prepared, and their permeability was studied.
|
|
|
Electro-optical sensor for the detection of extracellular ions
Bellan, Boris ; Salyk, Ota (referee) ; Vala, Martin (advisor)
The aim of this work was to prepare a platform based on electrochemical transistor that would allow both electrical and optical detection of ion exchange between the electrolyte and the layer of organic semiconductor. Semiconductor that have been used in this work was poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate). The main goal was to verify whether the proposed platform meets the requirements of both characterization methods, and whether the sensitivity in the given arrangement is sufficient for the required range of analyte concentrations. The work also dealt with the study of the stability of the prepared platform in selected solvents and culture media, at the same time the method of optimizing the preparation of the photoresist foil and the completion of the OECT measuring cell was studied. A suitable solvent for polymethyl methacrylate was sought so that it did not affect the surrounding parts of the platform. Based on the obtained results, it was found that the selected photoresist for the preparation of the spacer frame is not suitable. However, it was found that the proposed platform satisfies both characterization methods. Therefore, it will be necessary to replace the current photoresist with a new, more chemically stable one.
|
|
|
Real-time monitoring of cellular processes - current approaches
Švecová, Iva ; Hodný, Zdeněk (advisor) ; Groušl, Tomáš (referee)
This thesis aims to provide an overview of real-time live-cell imaging methods with a focus on the signalling pathways. The first, most thorough section is about fluorescence methods and is followed by sections about bioluminescence and label-free methods. In the fluorescence section, we will at first introduce the types of fluorophores and respective labelling approaches. Subsequently, we will go through the individual techniques, starting with single-fluorophore and FRET biosensors, continuing with kinetic modelling approaches, a FLIM method used to detect changes in the cellular environment, and ending with two methods used to improve the resolution. With each technique, we will shortly explain the working principle and look at the examples at which this method was used. Finally, we will look at the example of live-cell imaging of one signalling cascade.
|
|
|
Elektrochemický biosensor pro deteci patogenních bakterií v potravinách
Jankovský, Viktor
The aim of the theoretical part of this work is to summarize vital pathogenic bacteria in the foodstuff, describe the essential characteristics of the chosen bacteria, the health consequences and the most-often used treatment. Next topic included in the theoretical part is treatise about not only the classical cultivational but also the modern technical methods used to detect bacteria. The last chapter discusses the essential electrochemical methods used in the practical experiment. The practical part is focused on the verification of the electrochemical biosensor functionality which is used for the detection of pathogenic bacteria. Special attention is paid to the cleaning of the gold electrodes, application of the thin thiol layer, activation of the surface, immobilization of the antibodies and bacteria and also to the electroche-mical-quantitative determination of the bacteria concentration. Considering that the bio-sensor construction represents a quite complicated experiment, it was necessary to em-phasize all the partial steps of the biosensor's construction so that it was possible to veri-fy the correctness of the procedure. The last step of the practical part is the analysis and interpretation of the results.
|
|
|
Study of possibilities of increasing the biocompatibility of organic semiconductor surfaces
Malečková, Romana ; Šafaříková, Eva (referee) ; Vala, Martin (advisor)
This bachelor thesis deals with the possibility of biocompatibilization of organic semiconducting polymer PEDOT:PSS using RGD peptide for the construction of biosensors. Samples were prepared and compared where the RGD peptide was bound directly, as well as samples with a peptide bound via crosslinker molecule sulfo-SANPAH. Whether the RGD peptide was bound to the substrate was determined by the method of measuring the contact angle of liquids with subsequent calculation of surface energy. The results were further verified by elemental analysis, infrared spectrometry and Raman spectroscopy.
|
|
|
Development of a sensing platform for the study of physiological functions of living cells
Marková, Aneta ; Víteček, Jan (referee) ; Vala, Martin (advisor)
The aim was to develop a sensing platform on the base of organic electrochemical transistor (OECT). The focus was on the preparation of proper electrode system and on optimalization of properties of thin layer of organic semiconductor. As a base, commercial glass substrates with integrated indium-tin oxide electrodes were chosen. Thin layers were prepared from organic semiconductor poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) by spin-coating. Four formulations of material were studied. Layers with different thickness were prepared and the dependence of transconductance on the thickness of the layer and ratio of width and length was observed. The degradation of electrode system was solved by galvanic plating with gold. Attention was also paid to modifications to PEDOT: PSS. It has been found that the optimal layer thickness for use in sensors is approximately 150 nm. By reducing the series resistance by using a silver paste, the transconductance of 23 mS was obtained for the Ink 2, for the Ink 3 the transconductance was 44 mS. Sensoric platforms with these transconductances can be used for detection of physiological functions of electrogenic cells, e.g. cardiomyocytes.
|
|
| |
|
|
Designing and testing of new metal nanosubstrates for biomolecular sensors based on surface-enhanced Raman scattering (SERS) spectroscopy
Peksa, Vlastimil ; Procházka, Marek (advisor) ; Matějka, Pavel (referee) ; Richter, Ivan (referee)
Title: Designing and testing of new metal nanosubstrates for biomolecular sensors based on surface-enhanced Raman scattering (SERS) spectroscopy Author: Vlastimil Peksa Department: Institute of Physics of Charles University Supervisor: doc. RNDr. Marek Procházka, Ph.D., Institute of Physics of Charles University Abstract: This experimental methodical work was aimed at the optimization of selected gold and silver substrates and their use in construction of SERS-based biosensors, including following practical application. Several types of substrates, fabricated via a combination of bottom-up techniques on solid surfaces, were tested. The properties of these substrates were examined with probe molecules, namely methylene blue, porphyrins and tryptophan, on a confocal Raman microspectrometer. Obtained findings about the influence of analyte application, objective focusing and internal intensity standard were exploited for optimization of measurement procedures with regard to sensitivity, accuracy and reproducibility. A method for quantitative detection of food dye azorubine (E 122) in commercially available drinks was developed, based on these findings. Its results have shown its potential as a pre-scan method for field application and preliminary testing. Keywords: Metal nanosubstrates, biomolecules,...
|
guest ::
