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.

Předložená práce se zabývá problematikou slepé filtrace obrazů z transmisního elektronového mikroskopu. V úvodu práce je uveden popis transmisního elektronového mikroskopu. Navazující část popisuje mechanismy interakce elektronů se zkoumaným vzorkem a z toho vyplývající zobrazovací techniky elektronové mikroskopie. Poslední kapitola teoretické části práce zahrnuje popis vybraných metod slepé filtrace obrazu zejména s využitím dekompozice obrazu na charakteristické složky. Taktéž je zde uveden výčet metod pro zhodnocení úspěšnosti filtrace. V praktické části jsou popsány aplikované metody slepé filtrace obrazů a výsledky filtrování. Jednotlivé metody jsou mezi sebou porovnány. Získané výsledky a využitelnost aplikovaných metod jsou zhodnoceny v diskuzi.

The aim of this diploma thesis is to describe the influence of individual operating parameters on thickness and temperature distribution of elastohydrodynamic contact under high slip ratio. In this work, the distribution of the lubricant film thickness is determined by optical interferometry and the temperature mapping in EHL contact is realized by infrared microscopy. Two optical filters were used to separate the radiation of contact surfaces and oil. Temperature of disk surface was measured using a sapphire disk with chromium layer that shielded radiation from ball and oil. The experimental device was modified to simulate the conditions of the various vectors between entrainment and sliding speed. The results extend existing information about behavior of EHL contact under high slip ratio conditions.

Diagnostic of photovoltaic cells defects is one of the key step in production. This dissertation thesis deals about diagnosis of photovoltaic cells by radiative recombination of electron - hole pairs. This radiative recombination is known as luminescence. Diagnostic methods using this local light emission in infrared region have innovative potential. The method of electroluminescence is one of the most widely used luminescence methods whose modifications are applied across the industry, especially in semiconductors. The main area of this thesis research is focused on non-destructive analysis of radiated infrared radiation of monocrystalline and polycrystalline solar cells in various modifications of standard electroluminescence method. In this research there are used two types of CCD cameras are used as luminescent radiation detectors. Method of photon emission microscopy is used for magnification element. Local light emission inspection by microscopy should have benefit to qualitative evaluation of solar cells defects. Radiation emitted by the solar cell has a wave character. This fact brings the potential of using some of the characteristics of the waves. We are talking about the intensity and mainly about new characterization by polarization during the detection of defects in the solar cell surface. Innovation of the measuring workplace of the standard electroluminescence method to use polarization analysis is one of the goals of this dissertation thesis.

Předložená dizertační práce s názvem “Rastrovací elektronová mikroskopie a její aplikace pro senzitivní vzorky” pojednává o problematice rastrovací elektronové mikroskopie v kontextu instrumentálního a metodologického vývoje vedoucího k inovativnímu řešení, které je dobře aplikovatelné zejména v mikrobiologickém výzkumu. Součástí práce je rozprava o historii a současném stavu elektronové mikroskopie (EM) jakožto vědecké zobrazovací a analytické techniky, tato část se nachází v úvodních kapitolách. Nepopiratelný přínos EM v biologických a lékařských oborech je dokazován mnoha citovanými vědeckými publikacemi. Předložená dizertační práce přináší novinky z oblasti přípravy preparátů a kryogenní rastrovací elektronové mikroskopie (cryo-SEM) vyvinuté na pracovišti Ústavu přístrojové techniky AV ČR, v.v.i. v Brně. Jedná se především o návrhy a výrobu speciálních držáků vzorků a vývoj nových metodik v oblasti přípravy mikrobiologických preparátů vedoucích k nalezení optimálních parametrů jednotlivých procesů. V experimentální části se nachází ověření metodologických postupů při studiu hydratovaných a na elektronový svazek senzitivních preparátů. Následné srovnání různých přístupů na definovaném biologickém systému z oblasti mikrobiologie přispívá k rozšíření interpretace doposud známých výsledků. Mezi zkoumanými mikrobiologickými kmeny byly biofilm-pozitivní bakterie Staphylococcus epidermidis a kvasinky jako Candida albicans a Candida parapsilosis, jež jsou považovány za klinicky významné, protože se podílejí na vzniku závažných infekcí zejména u imunokompromitovaných pacientů. Dále byl studován vliv růstu biofilmu bakterie Bacillus subtilis na biodeteriorizaci a biodegradaci poly--kaprolaktonových fólií. Vývoj v oblasti cryo-SEM byl aplikován ve výzkumu mikrobů s biotechnologickým potenciálem, jako jsou např. Cupriavidus necator a Sporobolomyces shibatanus.

This thesis deals with the problem of practical application of FDTD technique for simulation of image formation in coherence controlled holographic microscope. Various ways for obtaining scattering matrices are explored in detail and the optimal technique based on a rigorous calculation of the far field is proposed. The scattering matrix, containing information about the observed sample, is subsequently used in analytic calculation of holographic signal; two levels of approximation of pupil function are also evaluated. The results are compared with a traditional approach based on Rytov approximation resulting in specification of the parameter domain where the approximation is applicable. Based on the simulations of the microscope, the dependence of axial resolution on apertures of the objective and the condenser and sensitivity of the signal to changes of refractive index of the sample is also studied.

This graduation thesis deals with the problem of voltage contrast in ESEM. The purpose of this work was to verify influence of used detectors in the dependence on conditions in specimen chamber on the size of voltage contrast. With the conditions in specimen chamber we understand pressure and working conditions of signal detection. We used power transistor as specimen.

The first part of the thesis deals with electron microscopy and physical model fluid flow. In the next part, a three-dimensional model is created in programme SolidWorks. It is the model of scintilating detector with aperture or screen with three hundred little apertures. These models are submitted to analysis focusing on investigation of the gas flow at different pressure in chamber of specimen ESEM using Cosmos FloWorks system. The results of the analysis of both apertures variants are compared with the request to minimize the pressure in the trajectory of the secondary electrons as well as in the chamber of specimen.

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.

The introductory part of this thesis is focused on the theoretical analysis of solved problems as examining the toxicity of naphthoquinones plumbagine and juglone, especially with regard to their allelopatic action. The next section is focuses on the plant stress, caused by the action of stress factors leading to their death. There is an experimental protocol and the possibilities of determination the effect of naphthoquinones on the plant model. Experimental section discusses changing growth parameters of the plant samples in different concentrations of the naphthoquinone. There is examined cell viability and changes in the synthesis of the secondary metabolites. Data obtained by using spectrophotometric and microscopic analysis are evaluated with STATISTICA software and statistical significance are plotted.