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Study of mesenchymal stem cell migration based on principles of chemotaxis
Pošustová, Veronika ; Skopalík, Josef (referee) ; Chmelíková, Larisa (advisor)
The purpose of this Master thesis is to verify migration of mesenchymal stem cells on the principle known as chemotaxis. First part of this study is focused on cell migration in order to explain the whole migration process. Next part describes various chemotaxis methods and selected studies dealing with clinical applications of mesenchymal stem cells in different medical and biomedical fields. The following step describes confocal microscopy, which is used for acquiring images of the cells. The experimental part is focused on cultivation of mesenchymal stem cells in a laboratory, which is necessary for cell vitality. Furthermore, there are designed two main experiments. Firstly there is a 2D experiment with adherent cells for chemotaxis using -Slide Chemotaxis. Secondly Transwell migration test is designed and executed. Finally, the acquired images from confocal microscope are used for image processing, which was done in Matlab R2020a programming environment. The result of this processing is evaluation of cell confluence and migration. In the end, experimental part of this study was optimized according to recommended studies. The results are summarized in the conclusion with proposal for improvements of those methods.
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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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Focus techniques of optical measurement of 3D features
Macháček, Jan ; Honec, Peter (referee) ; Janáková, Ilona (advisor)
This thesis deals with optical distance measurement and 3D scene measurement using focusing techniques with focus on confocal microscopy, depth from focus and depth from defocus. Theoretical part of the thesis is about different approaches to depth map generation and also about micro image defocusing technique for measuring refractive index of transparent materials. Then the camera calibration for focused techniques is described. In the next part of the thesis is described experimentally verification of depth from focus and depth from defocus techniques. For the first technique are shown results of depth map generation and for the second technique is shown comparison between measured distance values and real distance values. Finally, the discussed techniques are compared and evaluated.
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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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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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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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ANALYSES OF STRUCTURE OF COLLAGEN FIBRES IN ARTERIAL WALL USING MODERN METHODS OF LIGHT MICROSCOPY
Turčanová, Michaela ; Kochová, Petra (referee) ; Hampl, Aleš (referee) ; Burša, Jiří (advisor)
The doctoral thesis deals with the analysis of the arrangement of collagen fibres in arteries and their correct evaluation and use in structurally motivated constitutive models of the material. The first part of the work is focused on the literature search of mechanical properties of arteries and on an overview of available methods for the detection of waviness, orientation and dispersion of fibres. Most works identify fibre angles as additional parameter from mechanical tests and thus degrade the structural nature of the model. The second part describes an automatic algorithm that can evaluate the local directions of fibres and their scattering from images from a polarizing microscope and structurebased hyperelastic constitutive models. Furthermore, there is an emphasis on choosing the most appropriate imaging method based on fluorescence microscopy, which will help us to distinguish the waviness and scattering of fibres. In the next part of the thesis, two experiments on porcine arteries are presented in order to determine the influence of different magnitudes of biaxial deformation on fiber orientation and dispersion. The last part of the work presents the evaluated structural parameters for porcine and human aortas, which were analyzed not only under polarized light, but also under a laser scanning confocal microscope, thanks to which the waviness of the fibers and their global direction were obtained.
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Analysis of adherent cells confluency in 2D culture
Bracková, Michaela ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
First part of this semester work dealing with the theoretical description of adherens cells, particularly structure and functions of this cells. Subsequently work is devoted to description of cell culturing, with mention of conditions which are necessary for cell culturing, following by substances which promote cell growth. Last part of theoretical research is concern with microscopy technique that is suitable for studying of adherent cells. Subsequently it is about fluorescence and confocal microscopes. Practical part dealing with cell culturing of adherent cells and the evaluation of realized experiments.
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