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Analysis of neurite directionality
Plišková, Diana ; Čmiel, Vratislav (referee) ; Odstrčilík, Jan (advisor)
Práca je zameraná na navrhnutie vhodnej metódy analýzy smerovosti neuritov. Využité boli snímky neurónov z fluorescenčnej mikroskopie. Pred samotnou segmentáciou bolo potrebné snímky predspracovať, pričom sa postupne využila úprava kontrastu, ostrenie a adaptívna filtrácia pomocou Weinerovského filtru. Jednotlivé návrhy metód segmentácie pozostávali z prostého prahovania, narastaním oblastí a využitím morfologických operácií. Následná analýza smerovosti využívala smer gradientov v obraze. Navrhnutá metóda bola využitá aj ako klasifikátor, ktorý dokázal rozdeliť jednotlivé snímky do skupín podľa smeru rastu.
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Multiparametric fluorescence spectroscopy
Lacko, Kata ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This diploma thesis deals with the possibilities of multiparametric fluorescence spectroscopy, since the main objective of this experiment was to evaluate the possibilities of multiparametric measurements in the fluorescence spectroscopy laboratory. A suitable fluorescence probe was proposed for this type of experiment that shows high sensitivity for pH changes in the environment, SNARF-4F AM, based on a literature research. The fluorophore was dissolved in solutions of different pH and this system was examined using a time-resolved spectrofluorimeter. The method named TRES (time-resolved emission spectra) was used to obtain the emission spectra of the probe and to find the emission maximum. Fluorescence intensity decay measurements as a function of wavelengths allowed to create deconvolution of the emission spectra, which provided information about the fluorescent lifetime and the relative representation of the states of probes in the solution. Later, the probe was dissolved in solutions of different density and pH - this system served for anisotropic measurements, during which the individual correlation-rotational times of the fluorophore were obtained. The obtained results were then used as the basis for multiparametric analysis, which was performed by using a fluorescence correlation microscope and a spectrograph. This combination allows to measure the necessary fluorescence parameters in one step. A standard operating procedure was created for the spectrograph’s control. On the basis of the obtained information the suitability, accuracy and sensitivity of the multiparametric analysis were qualified.
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Bigels - Preparation and Characterization
Mušková, Alexandra ; Krouská, Jitka (referee) ; Mravec, Filip (advisor)
This bachelor thesis deals with study of bigels, which are composed of hydrogel and oleogel. The aim of this work is to prepare and characterize bigels. Preparation of hydrogels was based on interaction between hyaluronan and cationic surfactant carbethopendecinium bromide. Oleogels were prepared by mixing a non-ionic surfactant (sorbitan monopalmitate) with sunflower oil. Individual bigels were prepared by mixing the various rations of hydrogel and oleogel, and were characterized using a fluorescence microscope and rheological measurements. Fluorescence observations were done on prepared samples using Nile Red, Perylene, HPTS, ATTO 655 and fluorescein. Rheological experiments show that pure oleogel is the strongest and most solid in comparison with bigels and hydrogels. The greater amount of oleogel in system is, the stronger bigel is.
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Combining the light of two sources in the integration sphere
Mellová, Katarína ; Šicner, Michal (referee) ; Dostál, Zbyněk (advisor)
For the development of the new generation of the controlled holographic microscope, is necessary to design a new fluorescence module. The work discusses the basic principles and application of fluorescence microscopy, including a description of the holographic microscope. Searching for available light sources used in fluorescence microscopy is performed and evaluated. The designed source combines radiation with selected excitation wavelengths using an integrating sphere, which also has a homogenizing effect. This innovative design ensures a quick and efficient change of excitation wavelengths due to the fluorophore used and the possibility of a direct connection of the source to the microscope. In order to choose a suitable arrangement for the source, three conceptual designs of its function are proposed. The selected designs were verified by simulation and suitable optical components were selected. The resulting construction is tested.
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Analysis of neurite directionality
Plišková, Diana ; Čmiel, Vratislav (referee) ; Odstrčilík, Jan (advisor)
Práca je zameraná na navrhnutie vhodnej metódy analýzy smerovosti neuritov. Využité boli snímky neurónov z fluorescenčnej mikroskopie. Pred samotnou segmentáciou bolo potrebné snímky predspracovať, pričom sa postupne využila úprava kontrastu, ostrenie a adaptívna filtrácia pomocou Weinerovského filtru. Jednotlivé návrhy metód segmentácie pozostávali z prostého prahovania, narastaním oblastí a využitím morfologických operácií. Následná analýza smerovosti využívala smer gradientov v obraze. Navrhnutá metóda bola využitá aj ako klasifikátor, ktorý dokázal rozdeliť jednotlivé snímky do skupín podľa smeru rastu.
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Bigels - Preparation and Characterization
Mušková, Alexandra ; Krouská, Jitka (referee) ; Mravec, Filip (advisor)
This bachelor thesis deals with study of bigels, which are composed of hydrogel and oleogel. The aim of this work is to prepare and characterize bigels. Preparation of hydrogels was based on interaction between hyaluronan and cationic surfactant carbethopendecinium bromide. Oleogels were prepared by mixing a non-ionic surfactant (sorbitan monopalmitate) with sunflower oil. Individual bigels were prepared by mixing the various rations of hydrogel and oleogel, and were characterized using a fluorescence microscope and rheological measurements. Fluorescence observations were done on prepared samples using Nile Red, Perylene, HPTS, ATTO 655 and fluorescein. Rheological experiments show that pure oleogel is the strongest and most solid in comparison with bigels and hydrogels. The greater amount of oleogel in system is, the stronger bigel is.
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Multiparametric fluorescence spectroscopy
Lacko, Kata ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This diploma thesis deals with the possibilities of multiparametric fluorescence spectroscopy, since the main objective of this experiment was to evaluate the possibilities of multiparametric measurements in the fluorescence spectroscopy laboratory. A suitable fluorescence probe was proposed for this type of experiment that shows high sensitivity for pH changes in the environment, SNARF-4F AM, based on a literature research. The fluorophore was dissolved in solutions of different pH and this system was examined using a time-resolved spectrofluorimeter. The method named TRES (time-resolved emission spectra) was used to obtain the emission spectra of the probe and to find the emission maximum. Fluorescence intensity decay measurements as a function of wavelengths allowed to create deconvolution of the emission spectra, which provided information about the fluorescent lifetime and the relative representation of the states of probes in the solution. Later, the probe was dissolved in solutions of different density and pH - this system served for anisotropic measurements, during which the individual correlation-rotational times of the fluorophore were obtained. The obtained results were then used as the basis for multiparametric analysis, which was performed by using a fluorescence correlation microscope and a spectrograph. This combination allows to measure the necessary fluorescence parameters in one step. A standard operating procedure was created for the spectrograph’s control. On the basis of the obtained information the suitability, accuracy and sensitivity of the multiparametric analysis were qualified.
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