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Imaging of fluorescently labelled mitochondria using Biplane_FPALM microscopy
Dostál, Marek ; Plášek, Jaromír (advisor) ; Valenta, Jan (referee)
Title: Imaging of fluorescently labelled mitochondria using Biplane FPALM microscopy Author: Bc. Marek Dostál Department: Fyzikální ústav UK Supervisor: prof. RNDr. Jaromír Plášek, CSc. Consultants: RNDr. Petr Ježek, DrSc. Mgr. Hana Engstová, Ph.D. Abstract: In this thesis the results are presented of a search for suitable PCFP markers for a visualization inner and outer mitochondrial membranes by unique biplane FPALM microscope. We participated in debugging the measuring software for the purpose of 3D vizualization mitochondrial nets and determine their parameters. For this purpose were developed two methods. The first one can be applied to determine the parametrs of outer membranes and the second one is applicable to determine parametrs of inner membranes. On smaller statistical file we managed to confirm dependence quality of mitochondrial net on the cell culture conditions. Under stress conditions e.g. hypoxia the quality of mitochondrial net is changed. Keywords: mitochondria, Biplane FPALM, GFP, 3D visualization.
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Biological imaging by super-resolution microscopy
Adamová, Zuzana ; Komrsková, Kateřina (advisor) ; Benda, Aleš (referee)
zuzana_adamova_abstract_eng.txt[11.05.2017 20:26:29] Fluorescence microscopy is one of the most widely used imaging techniques in biological research. Despite its numerous advantages, it can be used only for studies of structures larger than 200 nanometres, due to diffraction limit caused by a wave nature of light. The value of 200 nanometres is the best reachable value of optical resolution, in other words, the smallest distance of two objects, which can be separately recognized by conventional optical systems. Up to the end of the 20th century it was therefore impossible to observe finer details of cells. However, recently several breakthrough imaging techniques, named super-resolution microscopy techniques, managed to bypass the diffraction limit and enabled biologists to study much more delicate structures, such as small organelles, virions, protein complexes or even particular proteins, while still using a visual light. This thesis introduces some selected super-resolution methods, explains briefly their principles and presents some of their applications in biology.
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Imaging of fluorescently labelled mitochondria using Biplane_FPALM microscopy
Dostál, Marek ; Plášek, Jaromír (advisor) ; Valenta, Jan (referee)
Title: Imaging of fluorescently labelled mitochondria using Biplane FPALM microscopy Author: Bc. Marek Dostál Department: Fyzikální ústav UK Supervisor: prof. RNDr. Jaromír Plášek, CSc. Consultants: RNDr. Petr Ježek, DrSc. Mgr. Hana Engstová, Ph.D. Abstract: In this thesis the results are presented of a search for suitable PCFP markers for a visualization inner and outer mitochondrial membranes by unique biplane FPALM microscope. We participated in debugging the measuring software for the purpose of 3D vizualization mitochondrial nets and determine their parameters. For this purpose were developed two methods. The first one can be applied to determine the parametrs of outer membranes and the second one is applicable to determine parametrs of inner membranes. On smaller statistical file we managed to confirm dependence quality of mitochondrial net on the cell culture conditions. Under stress conditions e.g. hypoxia the quality of mitochondrial net is changed. Keywords: mitochondria, Biplane FPALM, GFP, 3D visualization.
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