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Fluorescence detection in pathogen analysis
Tomečková, Kristýna ; Kristýna, Zemánková (referee) ; Bezděková, Jaroslava (advisor)
This bachelor thesis deals with isolation and detection of bacteria. The used method is called molecular imprinting and is connected with fluorescence microscopy. The theoretical part concentrates on comparison of the developed method with methods that have been used till now. The practical part describes preparation and optimization molecularly imprinted polymers. These polymers were prepared on two different carriers – multititration wellplate and magnetic particles. The bacteria used as imprinted template was Enterococcus faecalis. Staphylococcus aureus was used as its competitor.
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Optimization of a method for the determination of inorganic ions using capillary electrophoresis
Špačková, Simona ; Bezděková, Jaroslava (referee) ; Vaculovičová, Markéta (advisor)
Inorganic ions are one of the most important group chemicals. Their importance is crucial for all living organisms on the planet and monitoring of their content is essential for this purpose in clinical or environmental analysis but also in the food industry, agriculture or many other industries. Many analytical methods for ion detection have been designed and used, depending on the area of their use. Recently, numerous methods have been adapted to areas where not primarily intended. One such methods may be, for example, laser ablation with inductively coupled plasma mass spectrometry, which has recently begun to affect the analysis of biological samples. As a complementary method, able to detect also non-metallic ions is capillary electrophoresis. In this master thesis, the possibility of applying capillary electrophoresis with indirect photometric detection for the separation of inorganic ions in cell lysates of HeLa cells by capillary electrophoresis was verified. Main attention was paid to chloride ions. The ability to separate inorganic compounds in cellular material using salicylic acid as an absorbing anion in the base electrolyte was tested.
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Use of nanomaterials to improve optical detection of biomolecules
Špačková, Simona ; Kristýna, Pavelicová (referee) ; Bezděková, Jaroslava (advisor)
The bachelor thesis deals with modern methods of biomolecule labeling. Attention is paid especially to luminescent labeling with semiconductor quantum dots, optimization of preparation and characterization of quantum dots (QDs) and their bioconjugates with Ala4 peptide, which has antibacterial properties. The first method of preparing QDs used UV-radiation and conjugation with the peptide of interest was carried out through the thiol group of the peptide. A second type of QDs was prepared thermally and conjugation with the peptide was performed by linkers EDC/sulfo-NHS, which allowed peptide bond formation between the carboxyl group of QDs and the amino group of the peptide. The next part of the thesis is focused on the influence of conjugation the antimicrobial activity of the peptide. To verify the antimicrobial properties of the peptide, an interaction between the QDs-Ala4 conjugate and Staphylococcus aureus was observed. Conjugates thus prepared were characterized by fluorescence spectroscopy and capillary electrophoresis with absorption detection (-214 nm).
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Optimization of a method for the determination of inorganic ions using capillary electrophoresis
Špačková, Simona ; Bezděková, Jaroslava (referee) ; Vaculovičová, Markéta (advisor)
Inorganic ions are one of the most important group chemicals. Their importance is crucial for all living organisms on the planet and monitoring of their content is essential for this purpose in clinical or environmental analysis but also in the food industry, agriculture or many other industries. Many analytical methods for ion detection have been designed and used, depending on the area of their use. Recently, numerous methods have been adapted to areas where not primarily intended. One such methods may be, for example, laser ablation with inductively coupled plasma mass spectrometry, which has recently begun to affect the analysis of biological samples. As a complementary method, able to detect also non-metallic ions is capillary electrophoresis. In this master thesis, the possibility of applying capillary electrophoresis with indirect photometric detection for the separation of inorganic ions in cell lysates of HeLa cells by capillary electrophoresis was verified. Main attention was paid to chloride ions. The ability to separate inorganic compounds in cellular material using salicylic acid as an absorbing anion in the base electrolyte was tested.
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Fluorescence detection in pathogen analysis
Tomečková, Kristýna ; Kristýna, Zemánková (referee) ; Bezděková, Jaroslava (advisor)
This bachelor thesis deals with isolation and detection of bacteria. The used method is called molecular imprinting and is connected with fluorescence microscopy. The theoretical part concentrates on comparison of the developed method with methods that have been used till now. The practical part describes preparation and optimization molecularly imprinted polymers. These polymers were prepared on two different carriers – multititration wellplate and magnetic particles. The bacteria used as imprinted template was Enterococcus faecalis. Staphylococcus aureus was used as its competitor.
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|
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Use of nanomaterials to improve optical detection of biomolecules
Špačková, Simona ; Kristýna, Pavelicová (referee) ; Bezděková, Jaroslava (advisor)
The bachelor thesis deals with modern methods of biomolecule labeling. Attention is paid especially to luminescent labeling with semiconductor quantum dots, optimization of preparation and characterization of quantum dots (QDs) and their bioconjugates with Ala4 peptide, which has antibacterial properties. The first method of preparing QDs used UV-radiation and conjugation with the peptide of interest was carried out through the thiol group of the peptide. A second type of QDs was prepared thermally and conjugation with the peptide was performed by linkers EDC/sulfo-NHS, which allowed peptide bond formation between the carboxyl group of QDs and the amino group of the peptide. The next part of the thesis is focused on the influence of conjugation the antimicrobial activity of the peptide. To verify the antimicrobial properties of the peptide, an interaction between the QDs-Ala4 conjugate and Staphylococcus aureus was observed. Conjugates thus prepared were characterized by fluorescence spectroscopy and capillary electrophoresis with absorption detection (-214 nm).
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