The development of micro and nanoelectronics and nanophotonics needs novel characterization techniques to ensure higher quality of designed devices. The thesis describes a use of Scanning Near-field Optical Microscopy (SNOM) in dimensional control and in local investigation of diverse physical parameters. As example of its potential, the correlation between object topography and reflection measurement of capacitors is shown.

First part of this thesis is a theoretical essay which deals with the basics of scanning electron microscopy, with structure and function of a scanning electron microscope, its’ special case of an various pressure scanning electron microscope, electron interaction with surrounding environment and with a scintillation detector. The applied part of the thesis is focused on evaluation of material contrast on Cu-W specimen. Material contrast is evaluated for different pressures of water vapors in the microscope specimen chamber and for different detection conditions.

The diploma thesis deals with the construction design of reflective type of holographic microscope together with the proposal for the mechanical imposition of optical elements. Thesis contains parts where the illumination beam tracing through the optical system is calculated and discussed, followed up with the accuracy requirements for rectification mechanisms and their design. The three-dimensional model of modularly solved microscope is presented in conclusion of this thesis together with selected microscope sub-assemblies.

Reflected-light digital holographic microscope developed at IPE FME BUT uses off-axis holography principle and low spatial and temporal coherence illumination. Microscope allows reconstruction of the image amplitude and the image phase, which can be handled in real time. The only limiting factors are imaging speed of the detector and computer performance when processing holograms. Reconstruction of image phase and amplitude allows high-resolution profilometric measurements in the vertical axis direction. This thesis deals with the automatization of profilometric measurement method proposed in [2]. Proposed method uses the combination of the image phase and the image amplitude for the measurement of specimens with surface structure the vertical size of which cause the uncertainty of the image phase by a factor of 2pí. Futher the thesis deals with the construction design of the illumination system of the microscope and its realization together with experimental verification of functionality of proposed method automatization.

The diploma thesis deals with theory of imaging in a transmitted-light digital holographic microscope using partially coherent illumination. The influence of spatial and temporal coherence state on optical sectioning property is solved. The coherent transfer function is calculated. From this function imaging characteristics for a two-dimensional scattering object are derived depending on its defocus. Two different designs of microscopes developed in the Laboratory of optical microscopy in IPE FME BUT are considered.

The aim of my Master's thesis is the design of a scanning electron microscope. The designed concept presents an innovative approach to the problems and respects all technical, ergonomic and aesthetical demands made on it. The main creative part of design process starting from the concept development and ending up with the final solution is preceded by a background research study including a historical, technical and design analysis of the electron microscope. The conclusion of my Master's thesis analyses the final design, its character and its contribution in broader context.

The perfusion chamber for long term observing of live cells by the means the Coherence-Controlled Holographic Microscope (CCHM) was designed. CCHM was built and designed at the Laboratory of the optical microscopy at the Institute of Physical Engineering, Brno University of Technology. CCHM can quantitatively evaluate dynamical changes inside live cells thanks to the quantitative information about phase shift in each pixel of the image. In order to demonstrate advantages of CCHM experimentally, it is important to keep the live cells in the good conditions. This is made by adding the fresh cultivation medium for studied cells directly in the microscope. In contrast to the stationary chamber the perfusion chamber allows both the cultivation medium exchange and the application of biological reagents without the necessity of removing the chamber from the microscope. Therefore we can study the vital signs of cells before and after the application of reagents. An original perfusion system with accessories compatible with CCHM was designed. The design is based on the previously published perfusion system solutions that are referred to in this thesis. The flow characteristics and medium exchange process was discussed and a modification of the internal geometry, based on numerical simulations, was introduced. The applicability of this perfusion chamber has been proven for the CCHM and even for different types of microscopes. The reactions of tumor and epithelial cells during the change of the environment from the cultivation medium to the physiologically solution were studied.

Práce se zabývá návrhem a implementací testovacího zařízení pro manipulačnínmechanismus vzorku v elektronovém mikroskopu. Testovací zařízení a jeho software zajištuje meření několika parametrů mechanismu, jejich statistické vyhodnocení a porovnání se specifikací. Cílem je vytvořit komplexní testovací zařízení s jednoduchým uživatelským rozhraním, s požadavkem náhrady stávajícího nemodulárního a nestabilního řešení a jeho rozšíření o testování dalších parametrů.

Diploma thesis deals with the design of a vibration isolated platform for low temperature scanning tunneling microscope working under ultra high vacuum (UHV STM). Cooling of the microscope is done by liquid helium using a flow cryostat designed in Institute of Scientific Instruments of the AS CR. In the thesis, general requirements of designing of an ultra high vacuum compatible devices are discussed, as well as the ways of vibrational isolation and damping. Also some ways how to restrict the transfer of vibration between vacuum devices and surroundings are mentioned. This knowledge is then applied to the design of the antivibrational microscope platform compatible with low temperature usage. For better understanding of vibrational transfer and damping, a real model of the designed platform is made and vibrational transfer characteristics are measured and compared with the theory.

Subject of this master's thesis is the observation of influence of external environment on the living cells with the use of multimodal holographic microscope. The theoretical part is summarising the development of the holographic microscopy at IPE FME BUT. The theoretical part also describes multimodal holographic microscope, which allows non-invasive observing of living cells. The thesis also covers construction of the microscope, basic working instructions and the hologram processing method. The main subject of the thesis is the research on the topic of chemotaxis and osmotic processes in the cells. Experiments were designed for the purpose of this thesis to cover topics mentioned above. The experimental part of the thesis deals with cultivation of the cells, preparation of the sample and observation chambers and processing of the data. This part later focuses directly on the laboratory measurements. In all experiments, cells K2 (full name LW13K2) were observed.