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Fluorescence probes to study the properties of hydrogel systems
Máčala, Jakub ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This bachelors thesis is focused on possibility of selected fluorescent probes for study of properties of hydrogel systems. Chosen probes are Prodan for polarity study, HPTS for measurement of pH of the probe environment and platinum complex [2,6-Bis(2-pyridyl)fenyl-C,N,N]chloroplatinum(II) for its unique fluorescence properties and potential of energy transfer studies in hydrogel systems. Spectral properties of fluorophores were defined in selected set of solvents with different polarity with steady state fluorescence spectroscopy method. Properties of HPTS probe were also tested in buffers with different pH. Subsequently, hydrogels based on hyaluronan-septonex and dextran-SDS with incorporated fluorescence probes were prepared. Steady state fluorescence measurement of fluorophores from hydrogel were made using fluorimeter optical fibres. It was found, that probes Prodan and HPTS are suitable for study of polarity and pH of hydrogels and probably they will be able to be used for study of hydrogel with other techniques, like fluorescence microscopy, however platinum complex is, due to the low fluorescence intensity and possible quenching by polar water environment, probably not suitable for other measurements.
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Stability of vesicular systems using fluorescence spectroscopy techniques
Máčala, Jakub ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This thesis is focused on possibility of studying stability and fusion of catanionic vesicles with Förster resonance energy transfer. The mainly used technique was Time-Correlated single photon counting. Firstly, excitation and emission spectra of chosen probes were measured and donor-acceptor pairs were suggested: 5-hexadecanoylaminofluorescein with Octadecyl Rhodamine B, Bodipy 493/503 with rhodamine or DiI, perylene with fluorescein, DiO with DiI. Then, time-resolved measurements of suggested pairs from environment of catanionic vesicles with different content of cholesterol were made in order to track the FRET associated with fusion of vesicles. It was found out, that it is not possible to use DiO as a donor because of it’s inefficient solubilisation into vesicles. It is also not possible for Bodipy to be used as a donor, because of it‘s excimer formation. In case of using fluorescein as a donor, it was found, that there is ongoing homo-fret between fluorescein molecules. Thanks to this, fusion was tracked by addition of unstained vesicles. It was also possible to track fusion in longer period of time. Also perylene-fluorescein pair was found to be capable of tracking the fusion, but with the exception of vesicles with content of cholesterol of 43 mol. %, tracking of fusion was possible only in short period of time.
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Stability of vesicular systems using fluorescence spectroscopy techniques
Máčala, Jakub ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This thesis is focused on possibility of studying stability and fusion of catanionic vesicles with Förster resonance energy transfer. The mainly used technique was Time-Correlated single photon counting. Firstly, excitation and emission spectra of chosen probes were measured and donor-acceptor pairs were suggested: 5-hexadecanoylaminofluorescein with Octadecyl Rhodamine B, Bodipy 493/503 with rhodamine or DiI, perylene with fluorescein, DiO with DiI. Then, time-resolved measurements of suggested pairs from environment of catanionic vesicles with different content of cholesterol were made in order to track the FRET associated with fusion of vesicles. It was found out, that it is not possible to use DiO as a donor because of it’s inefficient solubilisation into vesicles. It is also not possible for Bodipy to be used as a donor, because of it‘s excimer formation. In case of using fluorescein as a donor, it was found, that there is ongoing homo-fret between fluorescein molecules. Thanks to this, fusion was tracked by addition of unstained vesicles. It was also possible to track fusion in longer period of time. Also perylene-fluorescein pair was found to be capable of tracking the fusion, but with the exception of vesicles with content of cholesterol of 43 mol. %, tracking of fusion was possible only in short period of time.
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Fluorescence probes to study the properties of hydrogel systems
Máčala, Jakub ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This bachelors thesis is focused on possibility of selected fluorescent probes for study of properties of hydrogel systems. Chosen probes are Prodan for polarity study, HPTS for measurement of pH of the probe environment and platinum complex [2,6-Bis(2-pyridyl)fenyl-C,N,N]chloroplatinum(II) for its unique fluorescence properties and potential of energy transfer studies in hydrogel systems. Spectral properties of fluorophores were defined in selected set of solvents with different polarity with steady state fluorescence spectroscopy method. Properties of HPTS probe were also tested in buffers with different pH. Subsequently, hydrogels based on hyaluronan-septonex and dextran-SDS with incorporated fluorescence probes were prepared. Steady state fluorescence measurement of fluorophores from hydrogel were made using fluorimeter optical fibres. It was found, that probes Prodan and HPTS are suitable for study of polarity and pH of hydrogels and probably they will be able to be used for study of hydrogel with other techniques, like fluorescence microscopy, however platinum complex is, due to the low fluorescence intensity and possible quenching by polar water environment, probably not suitable for other measurements.
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