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Biomarker detection using electrochemical method with microfluidic chip
Klepáčová, Ivana ; Svoboda, Ondřej (referee) ; Neužil, Pavel (advisor)
The thesis is focused on the development of the electrochemical system with microfluidic platform for the detection of multiple biomarkers. It analyses the use of biomarkers for the early diagnosis of cancer. The theoretical part contains basic information about voltammetric methods and microfluidic systems. The practical part provides solutions to the microfluidic chips, including the description of the used materials, designs, methodologies of preparation and conclusions from the testing of the manufactured microfluidic systems. The thesis describes the lock-in electrochemical system which measures the response of 4 electrochemical cells simultaneously. For the electrochemical system measurements, an electrochemical chip consisting of 64 electrochemical cells was used. The results of the analysis include the processing of the system tests and detected voltammetric curves of the Fe2+/Fe3+ solution and cysteine.
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Design of elongated right angle milling head for horizontal boring machine
Svoboda, Ondřej ; Tůma, Jiří (referee) ; Holub, Michal (advisor)
This thesis deals with design of right-angled elongated milling head for horizontal boring machine of FERMAT production. The thesis includes a detailed research of milling heads, a description of the head´s basic components and certain commercially available head´s comparison. Design part is describe components description and the design. Calculations are focused on determinating of forces generated during machining, setting of loading spectrum and the power load calculating individual gears during machining. In the next part is calculation of the reactions in the spindle bearings included, the equivalent load and durability of spindle bearings, spindle deformation and spindle bearings optimum distance. The thesis also includes spindle strength calculation, horizontal and vertical shafts analysis, gears design with helical teeth and gear strength. The thesis describes the design of extended milling head and particular design nodes which is supplemented by the 3D model. In this thesis is also included a drawing of assembly and components.
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Plasmide DNA isolation from bacteria and transfection to HEK293 cell line
Měsíčková, Klára ; Fohlerová, Zdenka (referee) ; Svoboda, Ondřej (advisor)
The isolation of plasmid DNA is an important and often used method in microbiology. The isolation itself is preceded by preparation of bacterial competent cells and by amplification of the plasmids. In this stage, plasmids CHR2, ASAP1, ASAP-3, ASAP-5 and Kir2.1. are first amplified in E.Coli bacteria of the DH5 strain and then isolated through the method of phenol-chloroform extraction. Gel electrophoresis and transfection to cellular line HEK293 are used for determining the correctness of the isolation.
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Cell layer cultivation in the microfluidic system
Kachan, Ksenia ; Svoboda, Ondřej (referee) ; Chmelíková, Larisa (advisor)
The theoretical part of this thesis describes the principles of cell culture in vitro and modern in vitro models for endothelial cell layer cultivation. It also describes the principles of confocal and fluorescence microscopy. The practical part is dedicated to working with cell cultures and realization of experiment with cell cultivation in microfluidic system. Cell layer was photographed using laser confocal scanning microscope Leica TCS SP8 X during the entire experiment. For processing and analysis of the obtained images, an algorithm in the software MATLAB was proposed.
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Network Protocols Semiautomatic Diagnostics
Svoboda, Ondřej ; Ryšavý, Ondřej (referee) ; Holkovič, Martin (advisor)
This thesis is about semiautomatic network protocol diagnostics and creating protocol description from eavesdropped communication. Several network eavesdropping techniques and some common programs for network analysis are introduced. Well-known network protocols are described, with focus on their communication messages. Some already existing methods for creating models from examples are mentioned and their characteristics defined. Next we design architecture of developed tool and some methods, that create protocol description. After that we explain implementation of this tool and finally the tool is tested and experimented with.
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Study of properties of voltage membrane sensor ASAP1 expressed in HEK293 cell line
Jablonská, Dominika ; Čmiel, Vratislav (referee) ; Svoboda, Ondřej (advisor)
This thesis deals with the problematice of measuring membrane potential and monitoring the propagation of electrical activity of cells. For this purpose, fluorescence membrane voltage sensors have been developed to detect changes in the membrane potential by changing their fluorescence intensity. The practical part is focused on the study of the properties of the ASAP1 fluorescence probe, which was transfected into the HEK293 cell line, which are kidney cells from the human embryo. Cell membrane potential was changed using the patch-clamp technique.
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