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Regenerace smyslových orgánů u raků
KOR, Golara
The antennae are a crucial sensory organ in crayfish that plays vital roles in their survival, communication, and orientation within their environment. They are equipped with sensory hairs that enable crayfish to detect various stimuli. Crayfish possess the unique ability to regenerate their antennae. Hemocytes are specialized cells within crayfish and play a crucial role in their immune system and contribute to the process of regeneration. We applied cryo-scanning and Transmission electron microscopies to study sensory organ morphology and regeneration in crayfish. In Chapter 2, cryo-scanning electron microscopy was used to identify potential differences in antennal morphology in six different crayfish species including marbled crayfish Procambarus virginalis, Mexican dwarf crayfish Cambarellus patzcuarensis, red swamp crayfish Procambarus clarkii, signal crayfish Pacifastacus leniusculus, common yabby Cherax destructor, and spiny-cheek crayfish Faxonius limosus. We observed significant differences among the six crayfish species in the ratios of antenna length, segment length and width to carapace length, and the number of segments. Examination of ultrastructural features unveiled differences in the distribution patterns of sensory hairs along the antenna and the morphology of the antennal surface. The varying morphology of antennae among studied species possibly indicates an adaptation to the specific conditions of their respective habitats. Furthermore, the outcomes demonstrated that a combination of variations in both antennal morphological characteristics and biometric measurements could effectively help us to distinguish the different studied crayfish species. In Chapter 3, the ultrastructural behaviour of hemocytes during coagulation and phagocytosis in the initial phases of injury in marbled crayfish was explored using transmission electron microscopy. During the coagulation process, hemocytes experienced marked transformations in morphology. The cytoplasmic granules exhibited a change from electron-dense to electron-lucent forms with the progress of coagulation. The transformed granules containing amorphous, electron-lucent material were observed to combine and release their contents into the extracellular space as part of the coagulation process. Additionally, it was observed that the nucleus contents also play a role in the coagulation process. Furthermore, the amputation of the leg led to substantial muscle degeneration, and phagocytic hemocytes start to take up the necrotic tissues. Besides, we observed the digested remains from phagocytized necrotic tissues merged into granules and other cellular components, thereby enhancing the granularity of the hemocytes and altering their cellular morphology. However, it is essential to note that hemocyte degranulation during coagulation could potentially reduce their granularity. Since morphological features are critical for classifying hemocytes, these morphological changes during coagulation and phagocytosis must be considered significant factors. The degenerated material that stored inside phagocytic hemocytes may have application in immunological and regeneration processes. In Chapter 4, we employed transmission electron microscopy to explore the ultrastructural aspects of potential immune cell involvement in nerve regeneration within crayfish antennae post-amputation. The findings indicated that, during nerve regeneration, all three forms of hemocytes were present. However, the granules within semi-granulocytes and granulocytes mainly contributed to the generation of new organelles such as mitochondria, the Golgi apparatus, and nerve fibers in the regenerated nerves of crayfish antennae. We revealed the transformation of granules of hemocytes into diverse organelles during the process of nerve regeneration at the ultrastructural level. In conclusion, these granules function as compact repositories of adaptable materials carried by immune cells, capable of transformatio
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Precise Image Registration Used for Correction of Geometrical Image Distortion
Zemčíková, Petra ; Odstrčilík, Jan (referee) ; Mézl, Martin (advisor)
Cílem předkládané diplomové práce je pomocí lícování obrazů přesně popsat distorzní pole pro následné odstranění geometrické distorze. Snímky zkreslené geometrickou distorzí pochází z prozařovacího elektronového mikroskopu. První část práce se zabývá zejména teorií spojenou s elektronovou mikroskopií, vznikem geometrické distorze a samotnou obrazovou registrací s důrazem na intenzitní flexibilní metody lícování. Ve druhé části je pak představena vytvořená metoda pro modelování geometrické distorze a lícování obrazů postižených slabou geometrickou distorzí. Vyvinutá metoda je následně otestována na testovacích i reálných datech a srovnána s existujícími popsanými metodami pro obrazovou registraci (například open-source softwarem Elastix).
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Image composition
Smital, Petr ; Petyovský, Petr (referee) ; Richter, Miloslav (advisor)
This thesis deals with methods of improving the quality of image output of an electron microscope by composing consecutive frames. The thesis contains a brief description of an electron microscope and the problems that deteriorate its image quality, and three basic mathematical algorithms that can be used to remove them. These are the simple averaging method, the method of least squares and the method of vector space. For all of the methods, the thesis describes ways of their implementation, the possibilities of their use and their mutual comparison. This thesis also deals with the problem of so-called superresolution, i.e. exceeding the physical resolution of a sensor using the methods mentioned above, and the problem of electron beam centring.
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Analysis of the impact of the conic aperture placement in the supersonic flow in the differentially pumped chamber
Rous, Miroslav ; Bílek, Michal (referee) ; Maxa, Jiří (advisor)
The thesis is focused on the medium dynamics simulation in the environmental scanning electron microscope. Specifically, it researches (examines) the conical aperture location effect in the differential pumped chamber and the width of the pumping channel in this chamber. The theoretical part deals with the environmental scanning microscope generally and other tools used in this thesis, e.g. ANSYS Fluen software, fluid turbulence, medium free path of molecules and electrons scattering. In the practical part, the work is focused on data processing from the ANSYS Fluent program and on their evaluation.
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Utilzation of physico-chemical analyses in study on microbial stress-response
Slaninová, Eva ; Pekař, Miloslav (referee) ; Sedláček, Petr (advisor)
This diploma thesis deals with the utilization of physico-chemical analysis in study on microbial stress–response. The main content of this work was to propose and optimize techniques and methods which are generally used in different industries. Two bacterial strains, Cupriavidus necator H16 and its mutant strain Cupriavidus necator PHB-4 producing polyhydroxyalkanoates (PHA) under certain conditions, were used as model microorganisms. Initially, microscopic teques such as transmission electron microscopy (TEM), cryo scanning electron microscopy (cryo SEM) and atomic force microscopy (AFM) were proposed and tested for the characteriozation of morphological differences of bacteria. Furthermore, viscoelastic properties of bacteria were determined and compared by the oscillatory tests of the rheology method considering another type of samples. Thermal analysis methods, differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) in particular, focused on water transport and behavior of intracellular water influenced by presence of the granules of PHB in bacteria. The last technique, utilized in the study, was dynamic and electrophoretic light dispersion during pH changes for characterization of the surface properties of bacteria such as size, zeta potential and the isoelectric point.
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Parasitic aberrations of the hexapole corrector of spherical aberration - analysis and corrections
Sopoušek, Jan ; Lencová, Bohumila (referee) ; Radlička, Tomáš (advisor)
Jednou z možností korekce sférické vady v elektronové mikroskopii je hexapólový korektor. Ačkoliv samotný princip korekce je poměrně dobře v literatuře popsán, jen relativně málo je věnováno samotnému seřízení hexapólového korektoru, jež je stěžejní pro správnou funkčnost. Práce je věnována analytickému rozboru vad seřízení a jejich vlivu na rozlišení obrazu za použití metody eikonálu a aberačních integrálů. Je ukázáno, že nejdůležitější roli v parazitických aberací hrají výchylky a náklon hexapólů. V závěru je pak popsáno, jakým způsobem je třeba hexapólový korektor seřídit pro odstranění parazitických vad.
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Analysis of the shape of the intake ports of the current concept differentially pumped chamber
Bílek, Michal ; Vyroubal, Petr (referee) ; Maxa, Jiří (advisor)
This master´s thesis discusses the analysis of the shape of the intake ports of differentially pumped chamber for the current concept. The first part is an introduction to the microscopy issue, then it is focused on ESEM microscopes and it also includes a describtion of the Flow dynamics and a mathematical describtion. In the following chapters the software Solid Works, where the model of differentially pumped chamber is created, and the software Ansys Fluent, which is used for analysis, are described. The second part concerns the proposed concepts and the results of their simulations. At the end of the work the results are analyzed and evaluated.
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Analysis of the gas flow when pumping vacuum for newly designed scintillation detector
Poruban, Milan ; Vaculík, Sebastian (referee) ; Maxa, Jiří (advisor)
The aim of this thesis is to study the issue of eniveromental scanning electron microscopy and pumping gas to create vacuum in the newly designed scintillation detector. Further, creating a model of recently proposed scintillation detector and simulating and analyzing pumping gas in differentially pumped chamber of detector. The theoretical part deals with electron microscopy, electron sources, electron optics and secondary electrons detectors. It is also presented which signals are generated by the electron beam on the surface of a solid. Further fluid flow issues and equations describing the flow in the solved chamber are dismantled. Furthermore, the impact of gaseous environment on the trajectory of primary electrons, because there are collisions of primary beam with atoms and molecules of gas. The following section discusses creating, quality and importance of the network in mathematical modelling. A method of a final volume used to calculate the differential equations describing the flow of gas at the premises of the detector is described . The practical part consists in creating a model of scintillation detector and analyzing the gas flow in drawing a vacuum in the newly designed scintillation detector. At the end the simulation results of gas flow are compared for different variants of apertures and various pressures on the neck of a scintillation detector designed for optimum performance of the detector. The outcome of this thesis is model of newly designed scintillation detector with optimized shapes of apertures according to functional requirements.
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Evaluation of gas flow in the scintillation detector at different profiles and sizes of holes in the screens that have shaped the network
Truhlář, Michal ; Špinka, Jiří (referee) ; Maxa, Jiří (advisor)
This work deals with the flow of gas in the scintillation detector. Thesis describes the function of an electron microscope, the composition, distribution and development of microscopes from the earliest to modern. The work also includes familiarization with SolidWorks 3D modeling in which is the electron microscope modeled and Cosmos FloWorks in which they are carried out simulations of gas flow. The paper is focused on the flow of gas in various shapes of the screen and various pressures in chamber of specimen. The results of the analysis of both apertures variants are compared with the request to minimize the pressures in the trajectory of the secondary electrons of well as in the chamber of specimen.
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