We are reporting on a study of the near-field sensitivity and resolution of a metal-dielectric probe (MDP). The propagation of the electromagnetic field across the probe was studied experimentally by means of time-domain terahertz spectroscopy and numerically simulated by CST MicroWave Studio 2008. Several localised areas at the probe end facet were distinguished and showed to be sensitive to the local dielectric properties and local anisotropy of the sample. Contrast and sensitivity measurements were conducted in several configurations of a MDP; the results were confirmed by simulations. The acquired data were analysed by using singular value decomposition that enabled separating independent physical phenomena in the measured datasets and filtering external disturbances out of the signal. Independent components corresponding to the changes in the output terahertz pulse upon varying the probe-sample distance and reflecting the local anisotropy in a ferroelectric barium titanate (BaTiO3) crystal were extracted and identified. The domain structure with characteristic dimensions of about 5 um was resolved during imaging experiments on the ferroelectric BaTiO3 sample, i.e. the resolved structures were ten times smaller than the characteristic dimensions of the end facet of the probe and forty times smaller than...

Das Ziel der vorliegenden Arbeit ist es, den Einsatz der konfokalen Laser-Scanning-Mikroskopie (LSM) für die Beurteilung von humanen Osteoblasten auf experimentellen Implantatoberflächen zu prüfen. Die acht Probekörper besaßen anodische Konversionsoberflächen, die aus den Elektrolyten Zirkoniumsulfat, Phosphorsäure und Kaliumfluorid sowie deren Kombinationen hergestellt waren. Als Positivstandard dienten TICER® und Cercon®, als Negativstandard commercial pure Titan (CPT/Reintitan). Diese Probekörper wurden in Chamber Slides mit einer humanen Osteoblastenkultur inkubiert. Am 3., 5., 7. und 10. Versuchstag erfolgte die immunhistochemische Darstellung von Bone-Sialoprotein (BSP). Anschließend wurden im LSM fluoreszenzmikroskopisch 3D-Bildstapel aufgenommen und mit dem 2D-mikroskopischen Verfahren verglichen. Es zeigte sich, dass das Bone-Sialoprotein einen Hinweis auf aktive Osteoblasten darstellt. Die BSP-exprimierenden Strukturen, die sich zu Volumina zusammenfügen, variieren an den verschiedenen Versuchstagen und bei den unterschiedlichen Oberflächenstrukturen. Eine deutlich erhöhte BSP-Expressionsrate wurde bei den elektrolytisch beschichteten Probekörpern aus Zirkoniumsulfat, Kaliumfluorid und Phosphorsäure erfasst. Ein eindeutiger Unterschied zwischen der 2D-Mikroskopie und der LSM ergab sich jedoch nicht. Die LSM erlaubt aber zusätzlich Aussagen über absorbierte Proteine an den Probekörperoberflächen durch eine um 90°-gedrehte Projektion der 3D-Bildstapel.

Microscopic techniques with high spatial and temporal resolution are required for in vivo studying biological cells and tissues. Adaptive lenses exhibit strong potential for fast motion-free axial scanning. However, they also lead to a degradation of the achievable resolution because of aberrations. This hurdle can be overcome by digital optical technologies. We present a novel High-and-Low-frequency (HiLo) 3D-microscope using structured illumination and an adaptive lens. Uniform illumination is used to obtain optical sectioning for the high-frequency (Hi) components of the image, and nonuniform illumination is needed to obtain optical sectioning for the low-frequency (Lo) components of the image. Nonuniform illumination is provided by a multimode fiber. It ensures robustness against optical aberrations of the adaptive lens. The depth-of-field of our microscope can be adjusted a-posteriori by computational optics. It enables to create flexible scans, which compensate for irregular axial measurement positions. The adaptive HiLo 3D-microscope provides an axial scanning range of 1 mm with an axial resolution of about 4 microns and sub-micron lateral resolution over the full scanning range. In result, volumetric measurements with high temporal and spatial resolution are provided. Demonstration measurements of zebrafish embryos with reporter gene-driven fluorescence in the thyroid gland are presented.

We present a HiLo microscope with an electrically tunable lens for high-contrast three-dimensional image acquisition. HiLo microscopy combines widefield and speckled illumination images to create optically sectioned images. Additionally, the depth-of-field is not fixed, but can be adjusted between widefield and confocal-like axial resolution. We incorporate an electrically tunable lens in the HiLo microscope for axial scanning, to obtain three-dimensional data without the need of moving neither the sample nor the objective. The used adaptive lens consists of a transparent polydimethylsiloxane (PDMS) membrane into which an annular piezo bending actuator is embedded. A transparent fluid is filled between the membrane and the glass substrate. When actuated, the piezo generates a pressure in the lens which deflects the membrane and thus changes the refractive power. This technique enables a large tuning range of the refractive power between 1/f = (-24 . . . 25) 1/m. As the NA of the adaptive lens is only about 0.05, a fixed high-NA lens is included in the setup to provide high resolution. In this contribution, the scan properties and capabilities of the tunable lens in the HiLo microscope are analyzed. Eventually, exemplary measurements are presented and discussed.

This thesis is focused on synthesis and polymerization of caprolactone and its derivatives by living ring opening polymerization (LROP), Self-assembly in aqueous solutions produced nanoaggregates comprised of amphiphilic block copolymers or telechelic polymers with incorporated boron compounds (phenyl boronic acids and boron clusters). Incorporation of boron compounds was facilitated either by covalent or non-covalent bonding. Obtained complex nanoparticle structures manifested stimuli-responsive behaviour and were investigated under varying conditions by combination of light scattering, fluorescence spectroscopy and electron microscopy. The obtained results on solution behaviour of polymers in combination with added value of boron compounds, yield general aspects of nano aggregate morphology, responsive character tuning and practical aspects of synthesis and self-assembly overcame in the preparation process. The publications wrote during this thesis are therefore adding valuable information to researchers engaged in biomedical utilization of such nano assemblies.

ALBA proteins were discovered in Archaea more than 30 years ago. They were gradually identified to be well conserved in Eucaryotes as well. A functional dimeric form of these proteins with DNA and RNA-binding capability was claimed in both mentioned domains of organisms. However, their roles diversified during evolution and vary in between organisms. In Archaea, ALBAs are involved in the genome organization and RNA-protein interactions. In Eukaryotes, there are presented two different subfamilies of ALBA proteins - Rpp20 and Rpp25 subfamily. A sole protein from each subfamily was identified in some organisms though they were multiplied in plants, respectively. These proteins can interact with each other and participate in ontogenetic development and stress responses. According to several studies, ALBA proteins were found to be involved in DNA stability maintenance or pre-rRNA splicing in the nucleus of Arabidopsis thaliana. However, they have been shown to play a role in the cellular metabolism and stress responses in cytoplasm. Six ALBA proteins were identified in the genome of A. thaliana, three from each subfamily. In this study, all heterodimeric protein- protein interactions were investigated by the bimolecular fluorescence complementation (BiFC) assay which revealed positive results in...

The main subject of this diploma thesis is the study of the effect of humidity on the total sterilization effect of the dielectric barrier discharge. Sterilization is a process which can eliminate all forms of life. The plasma sterilization is one of the methods that are suitable for sterilization of temperature and chemical sensitive materials. This sterilization method was proved to be effective on the wide spectrum of procaryotic and eucaryotic microorganisms. Basically, the main inactivation factors for cells exposed to plasma are heat, UV radiation and various reactive species. Dielectric barrier discharge (DBD) operating at atmospheric pressure was used for the sterilization of the samples. The discharge was generated in dry air and in humid air. The plasma power densities were 2 160 mW.cm-3, 2 279 mW.cm-3 and 2 760 mW.cm-3 (dry air) or 2 326 mW.cm-3 and 2 850 mW.cm-3 (humid air). Humidity of air was achieved using a wash bottle filled with water through which air flowed into the DBD reactor. Fungi spores of Aspergillus niger were used as model microorganisms. Whatman paper No. 1 was used as the carrying medium. When comparing sterilization efficiency of humid and dry air operating at the same conditions, the higher sterilization effect was observed in humid air. The sterilization effect of the DBD generated in air was compared with results obtained during plasma generation in argon and nitrogen. At the same conditions, the highest sterilization effect was observed in argon, followed by humid air, nitrogen and dry air. It was found out that in our experimental setup the active species are probably the main inactivation mechanism. The influence of temperature on the inactivation of microorganisms was completely negligible. The discharge parameters were studied by means of the optical emission spectroscopy (OES). Plasma treated samples were analyzed employing scanning electron microscopy (SEM). Damage of the microorganisms due to the effect of plasma as well as plasma effect on the structure of the carrying medium was evaluated.

This work deals with tool steel Vanadis 6. It describes the preparation of samples for the three-point bending test, with which it examines the influence of surface roughness, the effect of nitriding, coating and duplex coatings on fracture behaviour of the steel. First, it examines bending strength and the total energy required to work of fracture. The work also includes mapping and measurement of surface roughness on different layers. This measurement is performed on a confocal microscope.

Electron microscopy has become an essential component in many scientific fields, in which it contributes to new discoveries. The microscopy itself is continually being developed and the limits, which seemed to be insurmountable, have been overcome. The instruments have become user friendlier and their mobility enables flexible practical use in the field. The subject of this work is the design of a scanning electron microscope, the calculation of a magnetic curcuit of an immersion objective combined with standard lens, the theoretical calculation of a microscope resolution and the design solution of the mechanical parts of the microscope with a sample manipulator. The thesis includes a description of the electron microscopy development summarizing it briefly from the very beginning up to now. It also deals with electron sources, especially the Shottky cathode, which is to be the main object observed by the proposed device. The work also contains a description of the calculation of the microscope resolution as a function of the current density distribution. Another interesting issue included in the theoretical part is the signal detection, a description of several types of detectors and possible signal processing. The solution of the thesis includes a description of the concept of the scanning electron microscope with an explanation of the distribution of combined lens functions. The optical diagram shows the arrangement of the electron optics system and the distibution of pressure in the chamber of the microscope. The theoretical calculation is devoted to the magnetic curcuit design of the objective and to the resolution of the microscope for a given extent of working distances. Two modifications of the lens were designed – a standard simple objective and a combination of the standard objective with the immersion magnetic one. The results of both modifications are given for the parameters to be compared. The combined objective was designed with the possibility of use in two modes, as a standard and immersion lens. The deflection system is also divided into two modes, as a single deflection for the standard lens and as a two-dimension deflection for the immersion lens. Detectors for secondary electrons (SE) and detectors for back scattered electrons (BSE) will be used for the signal detection. The design of the microscope is another large part, which gives details on the most significant components of the microscope. The content of the technical solution is a three-dimensional computer model, created in Autodesk Inventor, which also includes a sample manipulator driven by piezoelectric actuators.

The thesis deals with the application of nondestructive testing methods (acoustic emission, electron microscopy, X-ray diffraction topography) to description and identification of stages in the fatigue process in aluminium alloys. The main contribution of this dissertation thesis consists of the approach and evaluation methodology of cyclic damage in the materials. It dealt especially with the verification of measured data by using data obtained using other nondestructive methods. The experimental results show that the field of nondestructive testing materials can fundamentally uncover the process of fatigue damage and improve current knowledge in this area. Combination of used methods has become a powerful tool for the identification and monitoring of the whole fatigue degradation process of materials.