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Study of cardiomyocytes viability using confocal microscopy
Chutný, Pavel ; Svoboda, Ondřej (referee) ; Čmiel, Vratislav (advisor)
This work describes the study of cardiomyocytes viability using confocal microscopy. The theoretic introduction covers the basic principle of fluorescence together with fluoresceins in cells biology. The struction of cardiomyocytes, their selection from the lively heart and the methods of viability examination is comprised in the next part. The third topic is about confocal mikroskopy and its posibilities in cardiomyocytes imaging. In next chapters are described the experiments and their evaluating.
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Study of cardiomyocytes viability using confocal microscopy
Chutný, Pavel ; Svoboda, Ondřej (referee) ; Čmiel, Vratislav (advisor)
This work describes the study of cardiomyocytes viability using confocal microscopy. The theoretic introduction covers the basic principle of fluorescence together with fluoresceins in cells biology. The struction of cardiomyocytes, their selection from the lively heart and the methods of viability examination is comprised in the next part. The third topic is about confocal mikroskopy and its posibilities in cardiomyocytes imaging. In next chapters are described the experiments and their evaluating.
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Viability evaluation on cultivated mezenchymal cells
Jehličková, Jana ; Odstrčilík, Jan (referee) ; Čmiel, Vratislav (advisor)
This thesis deals with the possibilities of cell viability determination and its evalution. The thesis in theory describes cell cultivation, different methods of staining cells and display options. In the practical part is mentioned the preparation of the observed experiment and there are shown the examples of cell images created by confocal microscopy. Additionaly there are created the data analysis algorithm in Matlab. For setting suitable analysis parameters is formed user interface.
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Measurement of Surface Texture Parameters of Products Using Contact and Non-contact Methods
Harčarík, Matej ; Hammer, Miloš (referee) ; Jankových, Róbert (advisor)
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.
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Photosynthetic cell suspension cultures quantitative image data processing
Vlachynská, Alžběta ; Búzová,, Diana (referee) ; Červený,, Jan (advisor)
This work was carried out in collaboration with the Department of Adaptive Biotechnologies, Global Change Research Centre AS CR. It deals with the quantitative analysis of photosynthetic cell cultures. It uses images captured by a confocal fluorescent microscope to the automatic determining the number of cells in the sample. The work consists of a theoretical analysis, which briefly describes fluorescence and confocal microscopy. It also concisely introduces a microscope Leica TCS SP8 X, which I used to scan data. One capture is devoted to the theory of digital image processing. The second part deskribes the development of algorithm for processing 3D data and simplified algorithm for processing 2D data and its program implementations in a programming environment MATLAB R2013b. Grafical user interface is explained in detail. Done measurement are presented at the conclusion. It mentions compiled sample preparation protocol. The results of the program are compared with manual counting. Number of cells per 1 ml are determined by created program in samples of cell cultures Chenopodium rubrum (Cr) and Solanum lycopersicum (To).
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Advanced methods for cardiac cells contour detection
Spíchalová, Barbora ; Čmiel, Vratislav (referee) ; Odstrčilík, Jan (advisor)
This thesis focuses on advanced methods of detecting contours of the cardiac cells and measuring their contraction. The theoretical section describes the types of confocal microscopes, which are used for capturing biological samples. The following chapter is devoted to the methods of cardiac cells segmentation, where we are introduced to the generally applied approaches. The most widely spread methods of segmentation are active contours and mathematical morphology, which are the crucial topics of this thesis. Thanks to the those methods we are able in the visual data to accurately detect required elements and measure their surface chnage in time. Acquired theoretical knowledge leads us to the practical realization of the methods in MATLAB.
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Software FLIM system with pulse white light laser in confocal microscopy
Grund, Pavel ; Odstrčilík, Jan (referee) ; Čmiel, Vratislav (advisor)
The theoretical part of this master's thesis is focused on research of confocal microscopy and FLIM method. There are a principles and types of confocal microscopy and the use of broad-spectrum laser as a basic light source of these microscopes. It gives what the FLIM method and its use not only in cell biology. The practical part thesis includes the acquisition of three sets of fluorescence intensity images with use of applications tunable pulsed laser, function TimeGate and detection of hybrid detectors. For practical elaboration of this thesis is in the software Fiji created a plugin, which is the source code in the Java programming language. The types of plugins and their uses are described in the third chapter of the thesis. This plugin including the graphical user interface in the form of the dialog box, proceses the fluorescence intensity images and creates a graphical representation of data showing the fluorescence lifetime, so called pseudocolor map.
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The genome size determination in sturgeons using 2-D a 3-D image cytometry.
SRP, Jiří
The genome size of evolutionary polyploid, neopolyploid and hybrid sturgeons is well known for its high variability. Aim of this study was 1) to specify the genome size of polyploid and neopolyploid sturgeons using an analysis of 2-D and 3-D images of specifically stained cells nuclei, 2) to evaluate the samples of populations for cytogenetic analysis needs and thereafter, 3) to compare both methods and record the data either for next research or for negative selection from the broodstock. This test has been done at the laboratory of molecular, cellular and quantitative genetics, Faculty of Fisheries and Protection of Waters USB in Vodňany using all sturgeons spawners and using samples obtained from some foreign fish farms which cooperated with the faculty. The samples included A. ruthenus, A. baerii, A. stellatus, A. gueldenstaedtii and Huso huso, intentionally bred hybrids of A. gueldenstaedtii (8n) x A. baerii (12n), A. baerii (8n) x A. ruthenus (4n), A. gueldenstaedtii (8n) x A. baerii (10n) a A. gueldenstaedtii (8n) x A. ruthenus (4n). As methods have been chosen image cytometry and confocal microscopy which use image digitalization and subsequently its computer analysis. The genome size was measured from the size of specifically stained nuclei of erythrocytes in specimens sampled. Result of this study was measuring the genome size in sturgeons under study using different methods, recording the obtained data, description of spatial conformation changes of cell nucleus with increasing ploidy level and deduction of impact to their physiology and comparing the methods between each other. The conclusion is necessity of another sturgeons genome size determination , choice of the best methods for more effective search and research of non-standard individuals and subsequently an examination of their physiological differences.
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