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Measurement of the distribution of an equivalent of cell dry mass using quantitative phase contrast of the CCHM
Kovářová, Klára ; Čolláková, Jana (referee) ; Křížová, Aneta (advisor)
Subject of this bachelor's thesis is to create an overview of conversions of phase profile to the values of dry mass and to realize experiments verifying the importance of dry mass. In the theoretical part, the history of the holographic microscopy and the development of the holographic microscopy on IPE FME BUT in Brno are summarized. The transmission holographic microscope is also described. This thesis covers its construction description and basic working instructions. The theoretical part also deals with the importance of image phase. Two approaches are presented for the interpretation of the phase delay and there is research on phase conversions to dry cell mass (according to H. G. Davies and M. H. F. Wilkins, G. Popescue, G. Dunn and D. Zicha). In the experimental part cultivation of cells, preparation of observation sample and processing of data are described. This part of thesis is also focused on laboratory experiments. Cells K2 (full name LW13K2) were observed in all experiments. First experiments deal with nutritional deprivation of cells and later with cell growth.
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Development of Biophysical Interpretation of Quantitative Phase Image Data
Křížová, Aneta ; Jákl, Petr (referee) ; Vomastek, Tomáš (referee) ; Chmelík, Radim (advisor)
This doctoral thesis deals with biophysical interpretation of quantitative phase imaging (QPI) gained with coherence-controlled holographic microscope (CCHM). In the first part methods evaluating information from QPI such as analysis of shape and dynamical characteristics of segmented objects as well as evaluation of the phase information itself are described. In addition, a method of dynamic phase differences (DPD) is designed to allow more detailed monitoring of cell mass translocations. All of these methods are used in biological applications. In an extensive study of various types of cell death, QPI information is compared with flow cytometry data, and preferably a combination of QPI and fluorescence microscopy is used. The DPD method is used to study mass translocations inside the cell during osmotic events. The simplified DPD method is applied to investigate the mechanism of tumor cell movement in collagen gels.
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Observation of external environment influence on living cells with holographic microscope
Kovářová, Klára ; Tolde,, Ondřej (referee) ; Křížová, Aneta (advisor)
Subject of this master's thesis is the observation of influence of external environment on the living cells with the use of multimodal holographic microscope. The theoretical part is summarising the development of the holographic microscopy at IPE FME BUT. The theoretical part also describes multimodal holographic microscope, which allows non-invasive observing of living cells. The thesis also covers construction of the microscope, basic working instructions and the hologram processing method. The main subject of the thesis is the research on the topic of chemotaxis and osmotic processes in the cells. Experiments were designed for the purpose of this thesis to cover topics mentioned above. The experimental part of the thesis deals with cultivation of the cells, preparation of the sample and observation chambers and processing of the data. This part later focuses directly on the laboratory measurements. In all experiments, cells K2 (full name LW13K2) were observed.
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Coherence-controlled holographic microscope in cell's life cycle research
Křížová, Aneta ; Kozubek,, Michal (referee) ; Uhlířová, Hana (advisor)
The goal of this diploma thesis was using of a coherence-controlled holographic microscope in cell’s life research. A brief history of interference microscopy and it’s applications in biology is described. Also other microscopy techniques routinely used for transparent objects imaging are mentioned and the biology of cell’s life cycle briefly explained. Characteristics describing the shape of a cell were proposed and tested with respect to identification of particular phases of its life cycle. The method of dynamic phase differences was modified in order to distinguish the internal motion of cell’s mass from the movement of the whole cell. Selected characteristics were used to evaluate observations carried out with the holographic microscope and the possibilities of their further applications were depicted. In conclusion, obtained findings were summarized and modifications of microscope construction as well as data-processing software were suggested.
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Methodology of Dictyostelium discoideum imaging with the transmission digital holographic microscope
Křížová, Aneta ; Ježek, Jan (referee) ; Uhlířová, Hana (advisor)
The task of this bachelor’s thesis was to image Dictyostelium discoideum with a transmission digital holographic microscope and compare this imaging with a commonly used microscopy technique. This bachelor's thesis is structured into a theoretical part and an experimental part. The theoretical part describes the setting of the transmission digital holographic microscope, basic operation and physical principles of imaging with this microscope. Further the process of image reconstruction and numerical process of phase reconstruction is illustrated. At the end of this part, basic biological characteristics of observed cells of Dictyostelium discoideum are introduced. The experimental part describes the cultivation and preparation of Dictyostelium discoideum for the observations, and further the process of performed experiments. The cells of Dictyostelium discoideum have been imaged in couple experiments by a method of the classical phase contrast. By the transmission digital holographic microscope the cells of Dictyostelium discoideum have been imaged in two mediums. Images captured by the transmission digital holographic microscope were processed by a method of Dynamic phase differences and analysed.
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Testing of multimodal holographic microscope in reflection mode
Kachtík, Lukáš ; Slabý, Tomáš (referee) ; Křížová, Aneta (advisor)
The thesis is focused on testing of reflection mode of Multimodal holographic microscope constructed in cooperation with TESCAN company. The assessment of axial and lateral resolution, verification of axial intensity response and documentation errors and deficiencies found while measuring are included in the testing procedure. In the conclusion, the Multimodal holographic microscope is compared to an atomic force microscop. A proper understanding of microscope operation and a formulation of an adjustment procedure were essential components of this work.
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Cell Tracking in Multimodal Holographic Microscope Using Transmission Mode
Blašťáková, Lucia ; Křížová, Aneta (referee) ; Štrbková, Lenka (advisor)
This thesis is focused on the monitoring of cell dynamics in multimodal holographic microscope using transmission mode. The goal of this experiment is a quantitative description of the epithelial-mesenchymal transition of cancer cells NMuMG. The introduction briefly describes a history of interference microscopy and its applications, design of multimodal holographic microscope Q-Phase and work with the program used for data processing. Next follows a brief description of parameters that were selected to quantify the epithelial-mesenchymal transition. In conclusion, all results from the observation of epithelial-mesenchymal transition were summarized.
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Observation of Influence of Thin Layers on Cell Behaviour by Multimodal Holographic Microscope
Vengh, Martin ; Křížová, Aneta (referee) ; Štrbková, Lenka (advisor)
Surface treatment of materials for the cell-surface interaction and imitating intrinsic environment of cells is still subject of matter for their potentional usage in the bioengineering. One of the options to modify the surface of material is exhibiting them to effects of plasma of electrical discharge in steam of cyclopropylamine. In this type of plasma, the low pressure plasma polymerization takes place, which creates thin amine-rich layer. There is an assumption that this type of layer is appropriate for cell adhesion and proliferation. As a superior technique for a label-free monitoring of the cell-surface interaction, multimodal holographic microscope (MHM) Q-Phase was exploited to determine the biocompatibility of materials. MHM enables observation in quantitative phase imaging, where the phase is directly proportional to the cell dry mass. This gives opportunity to define various features able to determine the biocompatibility of materials. According to the results amine-rich films enhanced the conditions for the cell adhesion and proliferation.
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Selected Musical Activities Usable in the School Club
KŘÍŽOVÁ, Aneta
The thesis entitled Selected Musical Activities Usable in the School Clubs deals with the school club as a place for leisure and musical activities that can be used here. In the introductory chapters of this theoretical thesis describes the basic characteristics of the target group, school club as a place for spending leisure time and at the same time as a facility for leisure education and music activities. In the next part of the thesis, I describe my own proposal for music activities school clubs, which can serve as an inspiration for teachers and other educators.
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Development of Biophysical Interpretation of Quantitative Phase Image Data
Křížová, Aneta ; Jákl, Petr (referee) ; Vomastek, Tomáš (referee) ; Chmelík, Radim (advisor)
This doctoral thesis deals with biophysical interpretation of quantitative phase imaging (QPI) gained with coherence-controlled holographic microscope (CCHM). In the first part methods evaluating information from QPI such as analysis of shape and dynamical characteristics of segmented objects as well as evaluation of the phase information itself are described. In addition, a method of dynamic phase differences (DPD) is designed to allow more detailed monitoring of cell mass translocations. All of these methods are used in biological applications. In an extensive study of various types of cell death, QPI information is compared with flow cytometry data, and preferably a combination of QPI and fluorescence microscopy is used. The DPD method is used to study mass translocations inside the cell during osmotic events. The simplified DPD method is applied to investigate the mechanism of tumor cell movement in collagen gels.
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