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Cationic fluorescence probes in a polyanion-surfactant system.
Mondek, Jakub ; Šmejkalová, Daniela (referee) ; Pekař, Miloslav (advisor)
In this bachelor thesis were studied properties of aqueous solutions of hyaluronan and celytrimethylamonnium bromid with help of acridine orange as fluorescence probe. At first, this thesis was focused on interactions of acridine orange and hyaluronan. Emision and excitation spectra of acridine orange were observed. Result of these measurement was bonding of acridine orange dimer to carboxylic group of hyaluronan. Next studied interaction was interaction of cetyltrimethyl amonium bromid with acridine orange – hyaluronan system. Result of these interaction was, that molecules of cationactive surfactant pushed away dimers of acridine orange and took place of dimer of acridine orange on carboxylic group of hyaluronan. Dimer of acridine orange was changed into monomers. The reason why this thesis was focused on study of aqueus solutions of hyaluronan with cationactive surfactant is benefit of hyaluronan – surfactant system in research of transport system for target distribution of drugs.
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Diffusion properties of oppositely charged organic molecules in solutions of hydrophilic polyelectrolyte
Rýcová, Eva ; Márová, Ivana (referee) ; Mravec, Filip (advisor)
This work is focused on physical interactions of negatively charged polymers with small ionogenic fluorescent molecules. Trying to verify the presence of these interactions using fluorescence correlation spectroscopy (FCS) and provides a comprehensive view of the problem. The aim of this work is to observe the effect of concentration on the diffusion properties. P/D ratio, where P represents number of polymer binding sites and D number of dye binding sites, was chosen for this issue. Hyaluronate, sodium chondroitin sulfate and sodium polystyrene sulfonace were used as polymers and Acridine Orange, and Rhodamine 6G were chosen as fluorescent probes. The reason why this experiment uses these probes, was the assumption, that the positive charge occuring on the fluorescent probe will lead to the electrostatic interaction with the negatively charged polymer. As a result, the bond between acridine orange and polyelectrolyte was not clearly demonstrated, but the interaction with Rhodamine 6G have been proved.
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Utilization of spectroscopy in study on stress-resistance of bacteria on the sigle-cell level
Köbölová, Klaudia ; Venerová, Tereza (referee) ; Sedláček, Petr (advisor)
This diploma thesis deals with the possibilities of stress resistance analysis of the Cupriavidus necator H16 and PHB-4 bacterial cells by spectroscopic methods and by testing the suitability of acridine orange as a viable dye. Based on research in literature, suitable analytical methods have been proposed, namely flow cytometer and fluorescence microscope. The first part of the experimental work was focused on the fluorescence microscope, which confirmed the basic character of acridine orange. Three stress factors, 50% and 70% ethanol, and acidic pH (pH = 1) were selected for viability monitoring. The bacteria fluoresced with green color after exposure to ethanol and red spots were found next to the cells, indicating their loss of integrity. In an acidic environment, the bacteria fluoresced red because of a partial DNA breakdown. The results were verified by the combination of propidium iodide with SYTO9 and the acridine orange suitability proved to be useful in this method. Image records were processed using image analysis. In the second part, acridine orange was used to monitor fluorescence using a flow cytometer. The result of the measurement was fluorescence expressed as histograms for individual channels, where fluorescence was characterized by median and mean intensity. By comparing the methods used, the acridine orange appears to be a more suitable fluorescent dye for the microscope than for a flow cytometer in which it was more difficult to obtain cell viability information. In the last part of the experimental work interesting photophysical properties of acridine orange were investigated.
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Utilization of spectroscopy in study on stress-resistance of bacteria on the sigle-cell level
Köbölová, Klaudia ; Venerová, Tereza (referee) ; Sedláček, Petr (advisor)
This diploma thesis deals with the possibilities of stress resistance analysis of the Cupriavidus necator H16 and PHB-4 bacterial cells by spectroscopic methods and by testing the suitability of acridine orange as a viable dye. Based on research in literature, suitable analytical methods have been proposed, namely flow cytometer and fluorescence microscope. The first part of the experimental work was focused on the fluorescence microscope, which confirmed the basic character of acridine orange. Three stress factors, 50% and 70% ethanol, and acidic pH (pH = 1) were selected for viability monitoring. The bacteria fluoresced with green color after exposure to ethanol and red spots were found next to the cells, indicating their loss of integrity. In an acidic environment, the bacteria fluoresced red because of a partial DNA breakdown. The results were verified by the combination of propidium iodide with SYTO9 and the acridine orange suitability proved to be useful in this method. Image records were processed using image analysis. In the second part, acridine orange was used to monitor fluorescence using a flow cytometer. The result of the measurement was fluorescence expressed as histograms for individual channels, where fluorescence was characterized by median and mean intensity. By comparing the methods used, the acridine orange appears to be a more suitable fluorescent dye for the microscope than for a flow cytometer in which it was more difficult to obtain cell viability information. In the last part of the experimental work interesting photophysical properties of acridine orange were investigated.
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|
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Diffusion properties of oppositely charged organic molecules in solutions of hydrophilic polyelectrolyte
Rýcová, Eva ; Márová, Ivana (referee) ; Mravec, Filip (advisor)
This work is focused on physical interactions of negatively charged polymers with small ionogenic fluorescent molecules. Trying to verify the presence of these interactions using fluorescence correlation spectroscopy (FCS) and provides a comprehensive view of the problem. The aim of this work is to observe the effect of concentration on the diffusion properties. P/D ratio, where P represents number of polymer binding sites and D number of dye binding sites, was chosen for this issue. Hyaluronate, sodium chondroitin sulfate and sodium polystyrene sulfonace were used as polymers and Acridine Orange, and Rhodamine 6G were chosen as fluorescent probes. The reason why this experiment uses these probes, was the assumption, that the positive charge occuring on the fluorescent probe will lead to the electrostatic interaction with the negatively charged polymer. As a result, the bond between acridine orange and polyelectrolyte was not clearly demonstrated, but the interaction with Rhodamine 6G have been proved.
|
|
|
Cationic fluorescence probes in a polyanion-surfactant system.
Mondek, Jakub ; Šmejkalová, Daniela (referee) ; Pekař, Miloslav (advisor)
In this bachelor thesis were studied properties of aqueous solutions of hyaluronan and celytrimethylamonnium bromid with help of acridine orange as fluorescence probe. At first, this thesis was focused on interactions of acridine orange and hyaluronan. Emision and excitation spectra of acridine orange were observed. Result of these measurement was bonding of acridine orange dimer to carboxylic group of hyaluronan. Next studied interaction was interaction of cetyltrimethyl amonium bromid with acridine orange – hyaluronan system. Result of these interaction was, that molecules of cationactive surfactant pushed away dimers of acridine orange and took place of dimer of acridine orange on carboxylic group of hyaluronan. Dimer of acridine orange was changed into monomers. The reason why this thesis was focused on study of aqueus solutions of hyaluronan with cationactive surfactant is benefit of hyaluronan – surfactant system in research of transport system for target distribution of drugs.
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