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Analysis of autofluorescence retinal images
Mosyurchak, Andriy ; Odstrčilík, Jan (referee) ; Kolář, Radim (advisor)
Autofluorescence retinal images are obtained with a confocal laser scanning ophthalmoscope, and used for the diagnostic of glaucoma. Glaucoma causes a gradual death of nerve cells and can cause blindness. Retina autofluorescence is caused by pigment lipofuscin, which causes cell damage. The aim of this work was to study methods suitable for segmentation of autofluorescence zones and method for tracking objects in an image. In this project was implemented algorithm of autofluorescence zone detection using method of region growing, designed and realized method for tracking autofluorescence regions.
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Intrinsic fluorescence of bacteria Cupriavidus necator
Marková, Kateřina ; Obruča, Stanislav (referee) ; Mravec, Filip (advisor)
This thesis focuses on autofluorescence of flavins in gram-negative bacteria Cupriavidus necator H16 and its mutant strain PHB-4. The main methods used were fluorescence microscopy and flow cytometry. To confirm the presence of flavins, excitation and emission spectra of the bacterial suspension were measured, which were compared with flavin standards. In the part of testing cells without stress response, the autofluorescence of bacteria in PBS buffer and cell suspensions stained with fluorescence probe BODIPY 493/503 was measured. The ratio of short fluorescence lifetime to long autofluorescence lifetime, and its dependence on fluorescence probe was compared with previous conditions. Autofluorescence of the supernatant was measured; it was found that the relative amplitude of long lifetime was multiple times higher than in the cell. In the part devoted to the stress response, this thesis was focused on the amount of dissolved oxygen in the production medium and the effect on bacterial autofluorescence. Then differently concentrated hydrogen peroxide was used, the best results were obtained from the concentration of 100 mM in media. For comparison a combination of hydrogen peroxide with ferro-ammonium sulphate was used, but there was no big difference. Sodium azide and antimycin A were selected as substances that directly influence on bacterial respiratory chain. Both compounds affected change in the ratio of the relative amplitudes, but the distribution of these lifetimes and the autofluorescence change over time was affected only by sodium azide.
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New diagnostic methods in bronchology
Votruba, Jiří ; Daneš, Jan (advisor) ; Marel, Miloslav (referee) ; Pešek, Miloš (referee)
The aim of this work has been the development and optimization of methods for early diagnosis of lung cancer, their utility and integration into daily practice. Firstly, we developed a device for measurement of endobronchial temperature (thermobronchoscopy) and found significant difference in endoluminal temperature above tumors and infiltrated lymph nodes compared to healthy regions. We further designed an appliance for near infrared spectroscopy of the bronchial mucosa and identified spectroscopic features useful for localization of solitary pulmonary nodule. The use of the appliance improved yield of endobronchial biopsy compared to endobronchial ultrasound. In the next part of the study, we describe further techniques for early diagnosis of lung cancer including endobronchial ultrasound, optical coherence tomography, confocal fluorescence microendoscopy, reflectance spectroscopy, autofluorescence bronchoscopy, fluorescence bronchoscopy, and narrow band imaging with concise introduction of our experience gained in several pilot projects. Next, we showed the utility of measurement of acetic acid in exhaled air as a promising biomarker for non-invasive identification of patients with symptomatic acid gastroesophageal reflux. Lastly, we demonstrated significant difference in radiation dose in HRCT...
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Intrinsic fluorescence of bacteria Cupriavidus necator
Marková, Kateřina ; Obruča, Stanislav (referee) ; Mravec, Filip (advisor)
This thesis focuses on autofluorescence of flavins in gram-negative bacteria Cupriavidus necator H16 and its mutant strain PHB-4. The main methods used were fluorescence microscopy and flow cytometry. To confirm the presence of flavins, excitation and emission spectra of the bacterial suspension were measured, which were compared with flavin standards. In the part of testing cells without stress response, the autofluorescence of bacteria in PBS buffer and cell suspensions stained with fluorescence probe BODIPY 493/503 was measured. The ratio of short fluorescence lifetime to long autofluorescence lifetime, and its dependence on fluorescence probe was compared with previous conditions. Autofluorescence of the supernatant was measured; it was found that the relative amplitude of long lifetime was multiple times higher than in the cell. In the part devoted to the stress response, this thesis was focused on the amount of dissolved oxygen in the production medium and the effect on bacterial autofluorescence. Then differently concentrated hydrogen peroxide was used, the best results were obtained from the concentration of 100 mM in media. For comparison a combination of hydrogen peroxide with ferro-ammonium sulphate was used, but there was no big difference. Sodium azide and antimycin A were selected as substances that directly influence on bacterial respiratory chain. Both compounds affected change in the ratio of the relative amplitudes, but the distribution of these lifetimes and the autofluorescence change over time was affected only by sodium azide.
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New diagnostic methods in bronchology
Votruba, Jiří ; Daneš, Jan (advisor) ; Marel, Miloslav (referee) ; Pešek, Miloš (referee)
The aim of this work has been the development and optimization of methods for early diagnosis of lung cancer, their utility and integration into daily practice. Firstly, we developed a device for measurement of endobronchial temperature (thermobronchoscopy) and found significant difference in endoluminal temperature above tumors and infiltrated lymph nodes compared to healthy regions. We further designed an appliance for near infrared spectroscopy of the bronchial mucosa and identified spectroscopic features useful for localization of solitary pulmonary nodule. The use of the appliance improved yield of endobronchial biopsy compared to endobronchial ultrasound. In the next part of the study, we describe further techniques for early diagnosis of lung cancer including endobronchial ultrasound, optical coherence tomography, confocal fluorescence microendoscopy, reflectance spectroscopy, autofluorescence bronchoscopy, fluorescence bronchoscopy, and narrow band imaging with concise introduction of our experience gained in several pilot projects. Next, we showed the utility of measurement of acetic acid in exhaled air as a promising biomarker for non-invasive identification of patients with symptomatic acid gastroesophageal reflux. Lastly, we demonstrated significant difference in radiation dose in HRCT...
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Analysis of autofluorescence retinal images
Mosyurchak, Andriy ; Odstrčilík, Jan (referee) ; Kolář, Radim (advisor)
Autofluorescence retinal images are obtained with a confocal laser scanning ophthalmoscope, and used for the diagnostic of glaucoma. Glaucoma causes a gradual death of nerve cells and can cause blindness. Retina autofluorescence is caused by pigment lipofuscin, which causes cell damage. The aim of this work was to study methods suitable for segmentation of autofluorescence zones and method for tracking objects in an image. In this project was implemented algorithm of autofluorescence zone detection using method of region growing, designed and realized method for tracking autofluorescence regions.
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Autofluorescence dřevokazné houby Fomes fomentarius
Žižka, Zdeněk ; Gabriel, Jiří
We observed the core of wood-rotting fungus Fomes fomentarius fruiting bodies in fluorescent microscopes. When excited with blue light, they featured thin-walled generative hyphae with septa and clamps, some of which emitted a strong yellow, yellow-green to yellow-red autofluorescence. In addition, we found yellow-red fluorescing thick-walled skeletal hyphae and strongly fluorescent thick-walled pith sets. On using green excitation, intensive red fluorescence was observed not in all these fruiting body components but also in the rest of the wood material
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