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Lactobacillus DNA analysis using real-time PCR and HRM analysis
Aksamitová, Dagmar ; Illková, Kateřina (referee) ; Trachtová, Štěpánka (advisor)
The rapid and accurate identification of the bacterium of the genus Lactobacillus, which are an important part of the normal gastrointestinal microflora and fermented dairy products are currently mainly used amplification methods. The aim of the study was to analyze the possibility of resolution of selected bacterial strains of the genus Lactobacillus, using the metod of polymerase chain reaction in the real time combined with high resolution melting curve analysis (qPCR HRM). It was tested five primers designed for qPCR-HRM analysis of lactic acid bacteria. The specificity of the primers was also verified simultaneously using bioinformatic analysis. On the basis of analysis of the DNA were selected as the most appropriate primers P1V1/P2V1, V3F/V3R and V6F/V6R. The suitability of the primers V3F/V3R and V6F/V6R was verified on a complex sample of food supplement from which the DNA was isolated using magnetic particles. The presence of bacteria of the genus Lactobacillus was performed using high resoluting melting analysis curves. The obtained results were in agreement with the information given by the manufacturer.
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Cell Viability Measurement
Prokopyeva, Elena ; Sekora, Jiří (referee) ; Čmiel, Vratislav (advisor)
This master's thesis deals with the ways of cell viability determination and its measuring. The first part of this thesis has a brief introduction into the theory of cell viability, also the methods of cell viability measurement and different types of fluorescent probes are described. The thesis further includes a brief description of the image processing methods, as well as the fluorescent microscope and applied filters are described. In the practical part the system implemented at a graphical programming environment LabVIEW used for the cell viability evaluation is described, functions of applied units are explained. The program is checked on model data.
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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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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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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.
|
|
|
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.
|
|
|
Lactobacillus DNA analysis using real-time PCR and HRM analysis
Aksamitová, Dagmar ; Illková, Kateřina (referee) ; Trachtová, Štěpánka (advisor)
The rapid and accurate identification of the bacterium of the genus Lactobacillus, which are an important part of the normal gastrointestinal microflora and fermented dairy products are currently mainly used amplification methods. The aim of the study was to analyze the possibility of resolution of selected bacterial strains of the genus Lactobacillus, using the metod of polymerase chain reaction in the real time combined with high resolution melting curve analysis (qPCR HRM). It was tested five primers designed for qPCR-HRM analysis of lactic acid bacteria. The specificity of the primers was also verified simultaneously using bioinformatic analysis. On the basis of analysis of the DNA were selected as the most appropriate primers P1V1/P2V1, V3F/V3R and V6F/V6R. The suitability of the primers V3F/V3R and V6F/V6R was verified on a complex sample of food supplement from which the DNA was isolated using magnetic particles. The presence of bacteria of the genus Lactobacillus was performed using high resoluting melting analysis curves. The obtained results were in agreement with the information given by the manufacturer.
|
|
|
Cell Viability Measurement
Prokopyeva, Elena ; Sekora, Jiří (referee) ; Čmiel, Vratislav (advisor)
This master's thesis deals with the ways of cell viability determination and its measuring. The first part of this thesis has a brief introduction into the theory of cell viability, also the methods of cell viability measurement and different types of fluorescent probes are described. The thesis further includes a brief description of the image processing methods, as well as the fluorescent microscope and applied filters are described. In the practical part the system implemented at a graphical programming environment LabVIEW used for the cell viability evaluation is described, functions of applied units are explained. The program is checked on model data.
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