|
|
Preparation of model membranes to study their interactions with biopolymers using fluorescence correlation spectroscopy
Adamcová, Zuzana ; Márová, Ivana (referee) ; Mravec, Filip (advisor)
This diploma thesis is focused on preparation and characterization of supported lipid bilayers as simplified models of cell membranes. The bilayers were prepared from source system of lecithin liposomes in phosphate buffer using the vesicle fusion method on a cover glass sufrace hydrophilized by plasma. Three fluorescent probes – Nile red, Oregon Green DHPE and DiO – were utilized to characterize diffusion within the bilayer using fluorescence correlation spectroscopy. For this purpose Z-scan FCS, which is a method developed specially for planar samples, was used. After the process of preparation and characterization of supported lipid bilayer was optimalized, interaction between this artificial membrane and solution of hyaluronic acid in phosphate buffer was studied. It was found out, that addition of this biopolymer causes slowing the diffusion of the fluorescent probe within the bilayer.
|
|
|
Interactions between hyaluronan and cetrimide
Adamcová, Zuzana ; Sedláček, Petr (referee) ; Mravec, Filip (advisor)
Interactions between hyaluronan and cationic Cetrimide were studied using fluorescence spectroscopy and turbidimetry in model physiological solution (0,15 M NaCl) and water. Results obtained in water showed strong electrostatic interactions before reaching CMC of the surfactant, which were screened in presence of salt due to high ionic strenght of the solution. Behavior of Cetrimide-hyaluronan systems was compared with the pure TTAB in equivalent system. It was found, that changes in Cetrimide-HA system occure at lower concentrations of the surfactant than in TTAB-HA system. This was probably caused by presence of small amount of CTAB in Cetrimide.
|
|
|
Preparation of model membranes to study their interactions with biopolymers using fluorescence correlation spectroscopy
Adamcová, Zuzana ; Márová, Ivana (referee) ; Mravec, Filip (advisor)
This diploma thesis is focused on preparation and characterization of supported lipid bilayers as simplified models of cell membranes. The bilayers were prepared from source system of lecithin liposomes in phosphate buffer using the vesicle fusion method on a cover glass sufrace hydrophilized by plasma. Three fluorescent probes – Nile red, Oregon Green DHPE and DiO – were utilized to characterize diffusion within the bilayer using fluorescence correlation spectroscopy. For this purpose Z-scan FCS, which is a method developed specially for planar samples, was used. After the process of preparation and characterization of supported lipid bilayer was optimalized, interaction between this artificial membrane and solution of hyaluronic acid in phosphate buffer was studied. It was found out, that addition of this biopolymer causes slowing the diffusion of the fluorescent probe within the bilayer.
|
|
|
Interactions between hyaluronan and cetrimide
Adamcová, Zuzana ; Sedláček, Petr (referee) ; Mravec, Filip (advisor)
Interactions between hyaluronan and cationic Cetrimide were studied using fluorescence spectroscopy and turbidimetry in model physiological solution (0,15 M NaCl) and water. Results obtained in water showed strong electrostatic interactions before reaching CMC of the surfactant, which were screened in presence of salt due to high ionic strenght of the solution. Behavior of Cetrimide-hyaluronan systems was compared with the pure TTAB in equivalent system. It was found, that changes in Cetrimide-HA system occure at lower concentrations of the surfactant than in TTAB-HA system. This was probably caused by presence of small amount of CTAB in Cetrimide.
|
guest ::
