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Microrheology with Fluorescence Correlation Spectroscopy
Kábrtová, Petra ; Sedláček, Petr (referee) ; Mondek, Jakub (advisor)
A comparison of three passive microrheological techniques was made with the emphasis on FCS. Fluorescently labelled and unlabelled polystyrene particles were used to probe a microrheological response of glycerol solutions, Mili-Q water and sodium hyaluronate solutions. In addition, for FCS technique an approximated equation for calculation of MSD values was derived and verified. It was found that FCS outmatches current microrheological techniques of DLS and video-based particle tracking by its ability to gain a broader data range including the area in which, until now, it was impossible to describe a microrheological behaviour of samples reliably.
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Study of interaction in biopolymer-surfactant system
Sovová, Šárka ; Mondek, Jakub (referee) ; Pilgrová, Tereza (advisor)
Interaction between biopolymer-surfactant and stability of arising complexes were investigated using dynamic light scattering method Stability of the system and interactions between native hyaluronan and cationic surfactant cetyltromethyl-amonium bromide in aqueous solution depending on preparation method were investigated. System was prepared either adding CTAB to hyaluronan causing sequential micelle’s formation, or adding hyaluronan to already formed micelles. It was found out that preparing method influences properties of arisen complexes. Molecular weight of hyaluronan has an inconsiderable effect on interaction against to concentration of hyaluronan, which influences interaction of component significantly.
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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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Time-Resolved Fluorescence in Research of Hyaluronan-Colloidal Systems Interactions
Mondek, Jakub ; Táborský, Petr (referee) ; Peter, Kapusta (referee) ; Pekař, Miloslav (advisor)
The aim of the doctoral thesis was to study advanced fluorescence techniques and its use in colloids or hyaluronan-surfactant systems and hydrogels based on hyaluronan, respectively. Steady-state and time-resolved fluorescence were used to study excited state proton transfer fluroescen probes in hyaluronan-surfactant systems to asses the influence of hyaluronan hydration to its interactions with oppositely charged surfactants. Firstly, different excited state proton transfer fluorescence probes were discussed to choose the most suitable candidate for next research. The influence of hyaluronan on inner environment of micells was determined based on the sensitivity of excited state proton transfer of chosen fluorescence probe 1-naphtol and, based on above mentioned experiments, the structure of hyaluronan hydration shell was discussed. The next part of doctoral thesis was focused on fluorescence lifetime correlation spectroscopy and on the development of method of nanorheology. Measured correlation functions were transformed to mean square displacement with developed MATLAB script. Firstly, the fluorescence method was compared with well described methods such as videomicrorheology and dynamic light scattering to asses the reliability of fluorescence correlation spectroscopy in microrheology. Secondly, nanorheology method was developed and its use in passive nanorheology of hyaluronan hydrogels was discussed. Based on mentioned experiments, the fluorescence correlation spectroscopy microrheology and nanorheology methods were optimized to use the methods in hydrogel research.
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Fluorescence correlation spectroscopy in hydrophilic colloids
Rýcová, Eva ; Mondek, Jakub (referee) ; Venerová, Tereza (advisor)
This work is focused on studying of hydrophilic colloids using fluorescence correlation spectroscopy (FCS). The gel systems were studied with several fluorescent probes, in 0,15 M sodium chloride (NaCl) consist of hyaluronan and surfactant cetyltrimethylammonium bromide (CTAB). Probe ATTO 655 was chosen for closer examination due to the suppression of triplet state. Every surrounding occuring in the gel systems was examine individually. Results showed that the measurement of gel systems by this method can be realized under certain conditions, the repeatability of the results was loaded with a small standard deviation. Extension of diffusion times caused the presence of micelles, triplet state was caused by a characteristic environment of the hydrogel. The probe also showed free diffusion motion in all environments. This diffusion motion was represented by the shortest diffusion time.
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Time-resolved fluorescence of system polymer-surfactant
Mondek, Jakub ; Čeppan, Michal (referee) ; Mravec, Filip (advisor)
In this diploma thesis was studied time-resolved fluorescence in polymer-surfactant system. At first aggregation numbers of cationic (cetyltrimethylammonium bromide), anionic (sodium dodecylsulfate) and nonionic (Triton X-100) surfactants were studied by steady-state and time-resolved fluorescence spectroscopy. These two methods were compared. Aggregation numbers by steady-state method were always lower than aggregation numbers measured by time-resolved method. Steady-state method of determination aggregation numbers is useless for surfactants with high aggregation number and for aerated samples. Addition of hyaluronan to surfactant system was studied. There was observed change in aggregation number after addition of hyaluronan and change in percentage of dynamic quenching after addition of hyaluronan. Hyaluronan affected aggregation number of cetyltrimethylammonium bromide and Triton X-100. Hyaluronan increased percentage of dynamic quenching in cetyltrimethylammonium bromide and in Triton X-100. Pyren in sodium dodecylsulfate was quenched by sphere of action with negligible percentage of dynamic quenching and addition of hyaluronan had no effect on quenching. As next goal of this thesis, the determination of the position of fluorescence probe pyrene in cetyltrimethylammonium bromide, sodium dodecylsulfate and Triton X-100 micelles was chosen. Position of pyrene changed with charge and structure of micelles. Next was studied how percentage of dynamic quenching by iodide compounds changes with different charge of micelle. In all cases majority of dynamic quenching was calculated.
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Excited-state proton transfer as a tool for characterization of coloid particles
Richterová, Veronika ; Pilgrová, Tereza (referee) ; Mondek, Jakub (advisor)
This bachelor thesis deals with the study of excited-state proton transfer in micellar systems. At first critical micelle concentration of surfactants CTAB, SDS and Triton X-100 was determined. Further the steady-state and time-resolved fluorescence of probes that undergo excited-state proton transfer with these surfactants was measured. As probes were chosen 1-naphthol, HPTS and 3HNA. Deprotonation rates of 1-napthol and HPTS and average lifetime of 3HNA were calculated from time-resolved measurement. Steady-state fluorescence was used for observation surfactants influence on excited-state proton transfer.
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Time-Resolved Fluorescence in Research of Hyaluronan-Colloidal Systems Interactions
Mondek, Jakub ; Táborský, Petr (referee) ; Peter, Kapusta (referee) ; Pekař, Miloslav (advisor)
The aim of the doctoral thesis was to study advanced fluorescence techniques and its use in colloids or hyaluronan-surfactant systems and hydrogels based on hyaluronan, respectively. Steady-state and time-resolved fluorescence were used to study excited state proton transfer fluroescen probes in hyaluronan-surfactant systems to asses the influence of hyaluronan hydration to its interactions with oppositely charged surfactants. Firstly, different excited state proton transfer fluorescence probes were discussed to choose the most suitable candidate for next research. The influence of hyaluronan on inner environment of micells was determined based on the sensitivity of excited state proton transfer of chosen fluorescence probe 1-naphtol and, based on above mentioned experiments, the structure of hyaluronan hydration shell was discussed. The next part of doctoral thesis was focused on fluorescence lifetime correlation spectroscopy and on the development of method of nanorheology. Measured correlation functions were transformed to mean square displacement with developed MATLAB script. Firstly, the fluorescence method was compared with well described methods such as videomicrorheology and dynamic light scattering to asses the reliability of fluorescence correlation spectroscopy in microrheology. Secondly, nanorheology method was developed and its use in passive nanorheology of hyaluronan hydrogels was discussed. Based on mentioned experiments, the fluorescence correlation spectroscopy microrheology and nanorheology methods were optimized to use the methods in hydrogel research.
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Excited-state proton transfer as a tool for characterization of coloid particles
Richterová, Veronika ; Pilgrová, Tereza (referee) ; Mondek, Jakub (advisor)
This bachelor thesis deals with the study of excited-state proton transfer in micellar systems. At first critical micelle concentration of surfactants CTAB, SDS and Triton X-100 was determined. Further the steady-state and time-resolved fluorescence of probes that undergo excited-state proton transfer with these surfactants was measured. As probes were chosen 1-naphthol, HPTS and 3HNA. Deprotonation rates of 1-napthol and HPTS and average lifetime of 3HNA were calculated from time-resolved measurement. Steady-state fluorescence was used for observation surfactants influence on excited-state proton transfer.
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Time-resolved fluorescence of system polymer-surfactant
Mondek, Jakub ; Čeppan, Michal (referee) ; Mravec, Filip (advisor)
In this diploma thesis was studied time-resolved fluorescence in polymer-surfactant system. At first aggregation numbers of cationic (cetyltrimethylammonium bromide), anionic (sodium dodecylsulfate) and nonionic (Triton X-100) surfactants were studied by steady-state and time-resolved fluorescence spectroscopy. These two methods were compared. Aggregation numbers by steady-state method were always lower than aggregation numbers measured by time-resolved method. Steady-state method of determination aggregation numbers is useless for surfactants with high aggregation number and for aerated samples. Addition of hyaluronan to surfactant system was studied. There was observed change in aggregation number after addition of hyaluronan and change in percentage of dynamic quenching after addition of hyaluronan. Hyaluronan affected aggregation number of cetyltrimethylammonium bromide and Triton X-100. Hyaluronan increased percentage of dynamic quenching in cetyltrimethylammonium bromide and in Triton X-100. Pyren in sodium dodecylsulfate was quenched by sphere of action with negligible percentage of dynamic quenching and addition of hyaluronan had no effect on quenching. As next goal of this thesis, the determination of the position of fluorescence probe pyrene in cetyltrimethylammonium bromide, sodium dodecylsulfate and Triton X-100 micelles was chosen. Position of pyrene changed with charge and structure of micelles. Next was studied how percentage of dynamic quenching by iodide compounds changes with different charge of micelle. In all cases majority of dynamic quenching was calculated.
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