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System for measurement of concentration of intracellular calcium in LabVIEW
Kormanovskaya, Elena ; Kolářová, Jana (referee) ; Čmiel, Vratislav (advisor)
The master’s thesis deals with the methods and system for measurement of concentration of intracellular calcium in LabVIEW. The thesis included theoretical and practical parts. The thesis covers the theory of fluorescence principles, fluorescence methods and indicators for measurement of concentration of intracellular calcium. The procedure of measurement by means of fluorescence microscope, camera and indicators is also mentioned. Literary solution is presented. Block diagram of system for measurement of concentration of intracellular calcium at a graphical programming environment LabVIEW is presented in practical part of master’s thesis.
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System for measurement of concentration of intracellular calcium in LabVIEW
Pochlopeň, Timotej ; Ronzhina, Marina (referee) ; Čmiel, Vratislav (advisor)
Master‘s thesis deals with the methods and system for measurement of concentration of intracellular calcium in LabVIEW. The thesis covers the theory of fluorescence principles, fluorescence methods and indicators for measurement of concentration of intracellular calcium. The procedure of measurement by means of fluorescence microscope, camera and indicators is also mentioned. The theory dealing with confocal microscopy is presented and two techniques are compared. Literary solution is presented. The practical part describes the program with a graphical user interface used for acquisition and analysis of data measuring dynamics of intracellular calcium concentration. The experimental part describes the preparation and measurement of cells and evaluation of experiments.
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Intracellular Calcium Measurement Methods
Číhal, Martin ; Ronzhina, Marina (referee) ; Čmiel, Vratislav (advisor)
This work aims to focus on the issue of intracellular calcium. Not only the theory dealing with the history of fluorescent indicators but also the types of fluorescent indicators and methods of measurements are summarized. The individual steps of measurements on the fluorescent microscope are mentioned as well. There is also created a program for evaluting Ca2+ concentrations in cells made in matlab.
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Utilization of spectroscopy in study on stress-resistance of bacteria on the sigle-cell level
Köbölová, Klaudia ; Venerová, Tereza (referee) ; Sedláček, Petr (advisor)
This diploma thesis deals with the possibilities of stress resistance analysis of the Cupriavidus necator H16 and PHB-4 bacterial cells by spectroscopic methods and by testing the suitability of acridine orange as a viable dye. Based on research in literature, suitable analytical methods have been proposed, namely flow cytometer and fluorescence microscope. The first part of the experimental work was focused on the fluorescence microscope, which confirmed the basic character of acridine orange. Three stress factors, 50% and 70% ethanol, and acidic pH (pH = 1) were selected for viability monitoring. The bacteria fluoresced with green color after exposure to ethanol and red spots were found next to the cells, indicating their loss of integrity. In an acidic environment, the bacteria fluoresced red because of a partial DNA breakdown. The results were verified by the combination of propidium iodide with SYTO9 and the acridine orange suitability proved to be useful in this method. Image records were processed using image analysis. In the second part, acridine orange was used to monitor fluorescence using a flow cytometer. The result of the measurement was fluorescence expressed as histograms for individual channels, where fluorescence was characterized by median and mean intensity. By comparing the methods used, the acridine orange appears to be a more suitable fluorescent dye for the microscope than for a flow cytometer in which it was more difficult to obtain cell viability information. In the last part of the experimental work interesting photophysical properties of acridine orange were investigated.
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Utilization of spectroscopy in study on stress-resistance of bacteria on the sigle-cell level
Köbölová, Klaudia ; Venerová, Tereza (referee) ; Sedláček, Petr (advisor)
This diploma thesis deals with the possibilities of stress resistance analysis of the Cupriavidus necator H16 and PHB-4 bacterial cells by spectroscopic methods and by testing the suitability of acridine orange as a viable dye. Based on research in literature, suitable analytical methods have been proposed, namely flow cytometer and fluorescence microscope. The first part of the experimental work was focused on the fluorescence microscope, which confirmed the basic character of acridine orange. Three stress factors, 50% and 70% ethanol, and acidic pH (pH = 1) were selected for viability monitoring. The bacteria fluoresced with green color after exposure to ethanol and red spots were found next to the cells, indicating their loss of integrity. In an acidic environment, the bacteria fluoresced red because of a partial DNA breakdown. The results were verified by the combination of propidium iodide with SYTO9 and the acridine orange suitability proved to be useful in this method. Image records were processed using image analysis. In the second part, acridine orange was used to monitor fluorescence using a flow cytometer. The result of the measurement was fluorescence expressed as histograms for individual channels, where fluorescence was characterized by median and mean intensity. By comparing the methods used, the acridine orange appears to be a more suitable fluorescent dye for the microscope than for a flow cytometer in which it was more difficult to obtain cell viability information. In the last part of the experimental work interesting photophysical properties of acridine orange were investigated.
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| |
|
|
Intracellular Calcium Measurement Methods
Číhal, Martin ; Ronzhina, Marina (referee) ; Čmiel, Vratislav (advisor)
This work aims to focus on the issue of intracellular calcium. Not only the theory dealing with the history of fluorescent indicators but also the types of fluorescent indicators and methods of measurements are summarized. The individual steps of measurements on the fluorescent microscope are mentioned as well. There is also created a program for evaluting Ca2+ concentrations in cells made in matlab.
|
|
|
System for measurement of concentration of intracellular calcium in LabVIEW
Pochlopeň, Timotej ; Ronzhina, Marina (referee) ; Čmiel, Vratislav (advisor)
Master‘s thesis deals with the methods and system for measurement of concentration of intracellular calcium in LabVIEW. The thesis covers the theory of fluorescence principles, fluorescence methods and indicators for measurement of concentration of intracellular calcium. The procedure of measurement by means of fluorescence microscope, camera and indicators is also mentioned. The theory dealing with confocal microscopy is presented and two techniques are compared. Literary solution is presented. The practical part describes the program with a graphical user interface used for acquisition and analysis of data measuring dynamics of intracellular calcium concentration. The experimental part describes the preparation and measurement of cells and evaluation of experiments.
|
|
|
System for measurement of concentration of intracellular calcium in LabVIEW
Kormanovskaya, Elena ; Kolářová, Jana (referee) ; Čmiel, Vratislav (advisor)
The master’s thesis deals with the methods and system for measurement of concentration of intracellular calcium in LabVIEW. The thesis included theoretical and practical parts. The thesis covers the theory of fluorescence principles, fluorescence methods and indicators for measurement of concentration of intracellular calcium. The procedure of measurement by means of fluorescence microscope, camera and indicators is also mentioned. Literary solution is presented. Block diagram of system for measurement of concentration of intracellular calcium at a graphical programming environment LabVIEW is presented in practical part of master’s thesis.
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