In this master’s thesis, a mechanical design of an in-column secondary electrons (SE) detector is presented. It is an ultravacuum compatible fibre-scintillation detector designed for use in an ultrahigh vacuum scanning electron microscope (UHV SEM). The designed in-column SE detector was manufactured and tested upon overcoming R&D challenges. The first section of this thesis deals with theoretical basis needed for understanding of functional principles of UHV SEM system and means of SE’s detection. In the second section, mechanical design of the in-column SE detector is described. The last section describes functionality of the designed detector. Effectiveness of light shielding of the detector was tested and the detective quantum efficiency was measured. Finally, images created by the designed in-column detector and an in-chamber SE detector were evaluated and compared.

This thesis deals with the influence of process parameters on the contamination level of powder materials produced by mechanical alloying (MA) technology. For this purpose austenitic stainless steel 316 L and equiatomic CoCrFeNi high-entropy alloy (HEA) were prepared by high-energy ball milling. Both materials were milled in argon and nitrogen atmospheres from 5 to 30 hours. Spark plasma sintering method (SPS) was then used for consolidation of chosen powder samples. Chemical analysis of contamination within MA was carried out using combustion analysers for determination of carbon, oxygen, and nitrogen contents after different lengths of milling. Also differences in chemical composition of powder and corresponding bulk samples were measured. The microstructure analysis using scanning electron microscopy (SEM) of both powder and bulk materials was executed with focus on oxide and carbide presence and dispersion. Increasing content of carbon with increasing milling time was observed across all measured samples. This contamination is attributed to using milling vial made of tool steel AISI D2 (containing 1,55 wt. % of carbon). Increase of carbon content within consolidation using SPS was also observed. Milling of specimens using N2 as milling atmosphere caused higher contamination level in both AISI 316 L and HEA compared to milling in argon.

This diploma thesis deals with the issue of motility analysis in leukemia cells. An accurate description of the cell movement and the detection of differences in motility under experimental conditions can be obtained by quantitative analysis of cell motility using time-lapse recording. The first part of this work describes various types of tumor cell migraton. The second part focuses on methods of analysis of cell motility in tissue culture using time-lapse recording, which include image acquisition and processing. Part of this chapter describes a coherence-controlled holographic microscope, which was used in the practical part and for which an insert was designed to ensure the exact and stable position of the individual chambers. The last part is focused on the research of leukemic cell motility, which is concluded by a discussion of the obtained results. The appendix contains a published study included acknowledgement to the author of this diploma thesis for participation in the project.

This work deals with elemental analysis of inorganic materials, that are presented by blast furnace slag and geopolymeric material containing fly-ashes. In the theoretical part there are explained principles of elemental analysis of inorganic materials. Ways of dissolution of samples, optical emission spectroscopy and electron microscopy with energy dispersive x-ray analysis are commented. In experimental part the ICP analysis of oxide standards is described. The standards were used for estimation of EDS-correction factors. Then follow the preparation, proving and use of standards in analysis of blast furnace slag. The work is finished by exact analysis of concentration profiles of elements in interface of phases in geopolymeric material.

The main goal of this work is to introduce with the basic technologies of manufacturing MEMS (Micro-electro-mechanical-systems). Further there is mentioned properties and manufacturing of CNT (Carbon nanotubes), used in manufacturing of capacitance pressure sensor.

This project deals with the theme of environmental scanning electron microscopy (EREM). This method allows the examination of insulators and wet specimens without pretreatment and modification like drying and metallization. The principle of this method consists in using higher pressure in a specimen chamber. The pressure is within the range of 100 – 200 Pa. However, the pressure in the specimen chamber restricts the signal detection interference. The objective of the work is to explore the possibility of interference in secondary electron route detection by way of electrostatic field. The electrostatic field was realized with the system consisting of four electrodes located in front of the scintillation detector. It should have interfered the secondary electron´s trajectory to the detector chamber. The optimization of voltage on the electrodes was made by simulation program called SIMION. The simulation results were experimentally verified with laboratory EREM.

The aim of this research is to explore the influence of magnetic field on early somatic embryos (ESEs) of spruce and pine. The magnetic field is gradual. This is important for the experiment, each cluster of ESE was exposed to magnetic field of different intensity. The resulting report of this experiment consists of the comparison between various methods of image analysis and the difference in growth of ESEs due to different exposure to magnetic field and surrounding temperature. Enclosed is a proposal of ESE size computing algorithm from a given image.

This work describes briefly electron microscopy issue and physical model of fluid flow. Then presented diploma thesis will be describing analysis and evaluation of the impact of supersonic flow on the pressure and density in the path of the primary electron beam in the differential pumping chamber. For analysis, evaluation and examination of the issue will be used CAD and CAE (Computer Aided Engineering) programs. At the same time it will be developed a methodology of calculation applicable to supersonic flow in the low pressure system at ANSYS CFX.

The master’s thesis deals with the methods and system for measurement of concentration of intracellular calcium in LabVIEW. The thesis included theoretical and practical parts. The thesis covers the theory of fluorescence principles, fluorescence methods and indicators for measurement of concentration of intracellular calcium. The procedure of measurement by means of fluorescence microscope, camera and indicators is also mentioned. Literary solution is presented. Block diagram of system for measurement of concentration of intracellular calcium at a graphical programming environment LabVIEW is presented in practical part of master’s thesis.

A programmable illuminating system (PIS) uses a commercial multimedia projector together with a suitable optical relay system in order to illuminate specimens under microscope with transmitted light. The theoretical part of the diploma thesis describes some methods used in the optical transmission microscopy. All of these methods employ physical masks placed in the condenser front focal plane. In the case of the traditional methods the masks are used to enhance contrast (e.g. dark-field illumination) and resolution (oblique illumination). One of the methods (a condenser with rotating aperture) provides the information about the three-dimensionality of the specimen. The next part of the thesis contains the theory and the basic classification of the illuminating systems used in the optical transmission microscopy. An optical and mechanical design of the optical relay system used for PIS is introduced. The experimental part shows the results made with two different PIS arrangements. It was shown on two different specimens that the PIS provides the same illumination as the classical methods. It was also proved that the PIS can simulate the rotating aperture in the condenser front focal plane and thus to give the information about the three-dimensionality of the specimen. Some new static and dynamic illuminating methods were introduced.