This master’s thesis includes an overview of profile method of surface texture evaluation according to ISO standards and VDA automotive standards, as well as an overview of areal method according to ISO 25178 standard series, which includes a proposal for improvement of czech terminology. Suitability of application of selected measurement methods for measurement of a portfolio of automotive parts was evaluated based on measurements carried out using a contact profiler, a coherence scanning interferometer and a confocal microscope. Further recommendations for practice in surface texture metrology were also formulated. A regression analysis of the relationship between values of selected profile surface texture parameters and their areal equivalents was also performed. Its results may aid the spread of areal surface texture evaluation in the industry.

The aim of this thesis is to study the issue of eniveromental scanning electron microscopy and pumping gas to create a 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 and the results compare with the previous model. 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 microscope 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. Furthermore, the simulation results are compared with the results of simulations on the older type of scintillation detector. The output of this thesis is model of recently proposed scintillation detector with visualized simulation results.

Dizertační práce se zabývá mechanismy mazání v náhradách kyčelního kloubu. Byla provedena systematická studie formování proteinového filmu při zahrnutí různých materiálů a provozních podmínek. Hlavní pozornost je přitom věnována roli jednotlivých proteinů obsažených v synoviální kapalině při současné přítomnosti dalších proteinů. Jelikož metody aplikované v předchozích studiích neumožňovaly separovat jednotlivé složky maziva, byla vyvinuta optická měřící metoda na principu fluorescenční mikroskopie. Z důvodu verifikace metody byly provedeny dvě nezávislé studie zaměřené na měření tloušťky mazacího filmu a dělení maziva na výstupu mazaného kontaktu. Z důvodu určitých limitací fluorescenční mikroskopie byla dále využita i metoda optické interferometrie, jejíž využití je ilustrováno ve studii zabývající se formováním mazacího filmu v náhradách kyčelního klubu při uvažování reálné konformity třecích povrchů. Závěrečná část práce představuje nový metodologický přístup založený na in situ pozorování kontaktní oblasti umožňující popsat roli jednotlivých proteinů ve vztahu k vývoji tloušťky mazacího filmu. Práce obsahuje originální výsledky, které přináší nové poznání v oblasti biotribologie náhrad kyčelního kloubu vedoucí k dalšímu vývoji implantátů při snaze zabránit jejich selhání v důsledku omezené životnosti.

This thesis deals with monitoring of the durability of FRP reinforcements in alkaline environments with differing temperature exposures. The theoretical part of the thesis consists of a general description of the FRP composites, the manufacturing of these FRP reinforcements using pultrusion technology, the resistance of these composites in aggressive alkaline environments, and the influence of temperature on the speed and manner of degradation. In the experimental part of this thesis, the FRP reinforcements were placed in an alkaline environment tempered at different temperatures (20 °C, 40 °C, 60 °C). Subsequently, any optical physical changes of the reinforcements were described and evaluated by the use of optical microscopy and the destructive tests of these reinforcements.

This diploma thesis deals with preparation and characterization of aqueous solutions of polyvinyl alcohol suitable for electrospinning. In the theoretical part method of electrospinning is described, including parameters which influence this process. Literary research also includes a chapter about rheology, which deals with the rheological properties of polymers, specifically PVA. The experimental part describes the preparation and rheological study of of aqueous solutions of polyvinyl alcohol, which were then spun. The morphology of prepared nanofibers was studied by using scanning electron microscopy (SEM). The effect of molecular weight, the solvent, concentration of solution, rheological properties, electrical conductivity and surface tension on the spinability, diameter and morphology of nanofibers is discussed.

This work is dealing with application of advanced fluorescence techniques for gaining knowledge about culture development during fermentation of red yeasts. Flow cytometry was used for auto-fluorescence measurement a carotenoids quantitation. It was resolved that while carotenoids are stored mainly in membranes the technique was feasible. If red yeast starts to accumulate carotenoids into lipid bodies mainly throughout the course of stationary phase, then the method starts to fail. Flow cytometric method using cell size measurement and light scatter for lipid quantitation was proved as applicable, too. However, it works only if cells are not starved. Individual calibration for each species is needed for elimination inter-species variations of intracellular structures. Fluorescence lifetime imaging microscopy was also used for studying of red yeast. Inherent ability to resolve different fluorescent species of the same molecule, which arise due to different molecular environment, helps with quantitation of cellular lipidic structures changes through the course of fermentation. Increase in the levels of carotenoids and/or rigidity of membranes was found as mechanism of protection during metabolic shifts, when intracellular content is vulnerable to damage.

V posledních letech se metody slepé dekonvoluce rozšířily do celé řady technických a vědních oborů zejména, když nejsou již limitovány výpočetně. Techniky zpracování signálu založené na slepé dekonvoluci slibují možnosti zlepšení kvality výsledků dosažených zobrazením pomocí elektronového mikroskopu. Hlavním úkolem této práce je formulování problému slepé dekonvoluce obrazů z elektronového mikroskopu a hledání vhodného řešení s jeho následnou implementací a porovnáním s dostupnou funkcí Matlab Image Processing Toolboxu. Úplným cílem je tedy vytvoření algoritmu korigujícícho vady vzniklé v procesu zobrazení v programovém prostředí Matlabu. Navržený přístup je založen na regularizačních technikách slepé dekonvoluce.

Background: Syllids are a species rich annelid family possessing remarkable regenerative ability, which is not only the response after traumatic injury, but also a key step during the life cycle of several syllid taxa. In these animals the posterior part of the body becomes an epitoke and is later detached as a distinct unit named stolon. Such a sexual reproductive mode is named schizogamy or stolonization. The prostomium and the proventricle, a modified foregut structure, have been proposed to have a control function during this process, though the concrete mechanisms behind it have never been elucidated. Results: By using different experimental set-ups, histology and immunohistochemistry combined with subsequent cLSM analyzes, we investigate and document the regeneration and stolonization in specimens of Typosyllis antoni that were amputated at different levels throughout the antero-posterior body axis. The removal of the anterior end including the proventricle implies an incomplete anterior regeneration as well as severe deviations from the usual reproductive pattern, i.e. accelerated stolonization, masculinization and the occurrence of aberrant stolons. The detailed anatomy of aberrant stolons is described. A histological study of the proventricle revealed no signs of glandular or secretory structures. The ventricle and the caeca are composed of glandular tissue but they are not involved in the reproductive and regenerative processes. Conclusions: As in other investigated syllids, the proventricle region has a significant role during stolonization and reproduction processes in Typosyllis antoni. When the proventricle region is absent, anterior and posterior regeneration are considerably deviated from the general patterns. However, proventricle ultrastructure does not show any glandular component, thereby questioning a direct involvement of this organ itself in the control of reproduction and regeneration. Our findings offer a comprehensive starting point for further studies of regeneration and reproductive control in syllids as well as annelids in general.

Diese Dissertation demonstriert die Nutzung von einzelnen optisch geheizten Goldnanopartikeln als Sensoren f ¨ur die Untersuchung von W¨armetransport und als Intensit¨atsmodulator f ¨ur Licht. Die beschriebenen Experimente basieren auf der photothermischen Mikroskopie, die die selektive Abbildung and Untersuchung von einzelnen absorbierenden Objekten erm¨oglicht. Goldnanopartikel werden optisch angeregt. Die Relaxation erfolgt durch nichtstrahlende Prozesse, die zu einer lokalen Erh¨ohung der Temperatur f ¨uhren. Die Erw¨armung f ¨uhrt zu einer Verringerung der Brechzahl, die als thermische Linse wirkt und dadurch die Ausbreitung eines zweiten nicht absorbierten Lichtstrahls vera¨ndert. Da die thermische A¨ nderung der Brechzahl sehr gering ist, wird das photothermische Signal durch das moduliertes Detektionsverfahren verst¨arkt. Der Heizlaserstrahl wird intensit¨atsmoduliert und erzeugt dadurch eine geringe Modulation der Strahlbreite des Detektionslaserstahls. Damit ver¨andert sich die Leistung, die durch eine Blende transmittiert wird. Diese Modulationsamplitude and Phaseverz¨ogerung werden mittels eines phasenempfindlichen Gleichrichters detektiert. Amplitude und Phase h¨angen von Modulationsfrequenz und thermischer Diffusivit¨at ab. Die frequenzaufgel¨oste Messung der beiden Gr¨oßen und deren Modellierung mittels einer verallgemeinerten Lorenz–Mie Theorie erm¨oglicht die Messung von der thermischen Diffusivit¨at des Mediums, das das Goldnanopartikel umgibt. In der zweiten Variante wird die Ausbreitung der W¨arme beobachtet. Ein Nanopartikel wird optisch geheizt und die ausgedehnte thermische Linse wird mit Hilfe der Ablenkung eines zweiten Laserstrahls vermessen. Das Ablenkungssignal wird mittels eines strahlenoptischen Models berechnet, um die thermische Diffusivit ¨at des Materials zu bestimmen, das das Nanopartikel umgibt. In einem weiteren Experiment wird das große Potential von optisch geheizten Nanopartikeln verdeutlicht. Einzelne Goldnanopartikel werden in eine d¨unne nematische Fl¨ussigkristallschicht eingebettet, deren Dicke darauf abgestimmt ist, dass die Schicht eine l/2-Platte darstellt. Die Goldnanopartikel werden optisch geheizt und steuern damit den Phasen¨ubergang von der nematischen zur isotropen Phase. Damit wird die Transmission eines zweiten Laserstrahls im Polarisationskontrast ge¨andert. Mit Hilfe dieser Anordnung kann die Intensit¨at eines Lichtstrahls um bis zu 100% moduliert werden. / This dissertation demonstrates the use of individual optically heated gold nanoparticles as sensors for investigations of heat transport and intensity modulation of light. The experiments employ the photothermal effect, which allows the selective detection and investigation of individual absorbers. The photothermal contrast is based on absorbing particles that are optically excited and relax via nonradiative processes. The absorbers act as nanosources of heat. The local temperature elevation leads to a local refractive index change due to thermal expansion which then acts as a lens. This thermal lens alters the propagation of a second non-absorbed beam of light. As the refractive index change with temperature is minuscule, the transmission changes of the detection are tiny as well. The photothermal signal is amplified by the use of a modulated detection scheme which enables the methods high sensitivity and provides a time scale for the measurement of thermal transport. The heating laser beam is intensity-modulated and thereby produces a small modulation of the beam waist of the detection laser beam and thus the transmitted power through an aperture. This modulation amplitude and phase are detected by a lock-in amplifier. Amplitude and phase depend on the modulation frequency and the thermal diffusivity of the material surrounding the nanoparticle. The frequency-resolved measurement of the two observables and their modeling using a generalized Lorenz–Mie theory allows the measurement of thermal diffusivities. In the second variant, the spread of heat into space is observed. A nanoparticle is optically heated, and the extended thermal lens is characterized by the deflection of a second laser beam. The deflection signal is modeled using ray optics to determine the thermal diffusivity of the material surrounding the nanoparticle. In a further experiment, the great potential of optically heated nanoparticles is demonstrated. Individual gold nanoparticles are embedded in a thin nematic liquid-crystal layer acting as a half-wave plate. The gold particles are optically heated. They control the transmission of a detection laser set up in polarization contrast. The intensity of the detection beam is modulated by up to 100%.

Diese Arbeit beschreibt die Untersuchung der Diffusion in dünnen Flüssigkeitsfilmen auf festen Oberflächen mit Methoden der Einzelmoleküldetektion. Anhand von Computersimulationen wird die Spotgrößenanalyse als neue Methode zur Analyse von Diffusionsprozessen vorgestellt. Die experimentellen Ergebnisse zeigen, dass die Diffusion von Farbstoffen in ultradünnen Flüssigkeitsfilmen stark verlangsamt ist. In einem ca. 3 nm dicken Film aus Tetrakis(2-ethyl-hexoxy)-silan werden Diffusionsprozesse beobachtet, die durch Sprünge zwischen Bereichen mit diskreten Werten der Diffusionskonstante gekennzeichnet sind. Die Existenz diskreter Werte der Diffusionskonstante kann durch die Ausbildung von Flüssigkeitsschichten molekularer Dicke (Liquid Layering) erklärt werden. Die Spotgrößenanalyse erweist sich als unabhängige Methode zur Bestimmung der Diffusionskonstante diffundierender Moleküle. Gegenüber der etablierten Bestimmung der Diffusionskonstante aus Trajektorien diffundierender Moleküle durch Berechnung der mittleren quadratischen Verschiebung bietet die Spotgrößenanalyse eine höhere statistische Genauigkeit und die Möglichkeit, Änderungen der Diffusionskonstante instantan zu detektieren.