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Microplastics in deep-sea fish
Fürychová, Barbora ; Cajthaml, Tomáš (advisor) ; Innemanová, Petra (referee)
Microplastics have become a significant problem in marine ecosystems, posing a serious threat to the biodiversity and stability of these environments. This bachelor's thesis focuses on investigating the presence of microplastics in the digestive tracts of deep-sea fish in the Indian Ocean. These fish represent interesting model organisms that can aid in understanding the distribution of microplastics in the marine water column. The introductory part of the paper summarizes the existing knowledge on microplastics, their occurrence, methods of analysis and the presence of microplastics in the aquatic environment. Particular attention is paid to deep- sea fishes, whose role in marine ecosystems is crucial. In the experimental part, detailed analyses of fish digestive tract samples were performed to detect and identify microplastics. The study confirmed the presence of microplastics in the digestive tracts of deep-sea fish, and various types of polymers were identified, including polystyrene, nylon, polyethylene terephthalate, polyvinyl chloride and polyester. This undergraduate thesis is the first study ever to demonstrate that microplastics are found in deep- sea fish in the western Indian Ocean. The conclusions of this thesis underscore the urgency of the situation regarding contamination of the...
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Filter wheel for confocal microscope
Daňková, Barbora ; Jákl, Petr (referee) ; Dostál, Zbyněk (advisor)
The existing holographic Q-Phase microscope, located at the FME of Brno BUT, is planned to be expanded with a confocal fluorescence module based on digital micromirror device technology. This will be implemented instead of the existing epi-fluorescence module. For the existing solution of the module, it is necessary to design a fluorescent filter turret, which was missing in it. This work explains the basic techniques of confocal fluorescence microscopy and deals in more detail with multi-channel fluorescence confocal microscopy principles. An optical simulation of its illumination part is created to realise the exchanger's design. Thanks to it, the dimensional requirements for the optical components of the turret are determined. Commercially available solutions are also evaluated. A custom solution is proposed since their parameters do not meet the requirements. This includes the mechanical design based on several concepts as well as its robotization. The turret is designed for six fluorescent channels, and drawing documentation has been compiled. According to her, the turret was manufactured, assembled, and successfully tested in the last part of the work.
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Concentration of grease constituents in the EHD contact and its surroundings
Palán, Miroslav ; Čermák, Jan (referee) ; Košťál, David (advisor)
The thesis focused on the study of the effect of thickener particle size and speed on the concentration of grease particles (thickener and base oil) in the contact and its close surroundings. The calibration images were taken on a sandwich module and the experiments were carried out in a ball-on-disk configuration. The distribution of particles in the grease was investigated by fluorescence microscopy, which allows separate observation of the individual particles. The results showed that the particle size of the thickener fundamentally affects its concentration in and around the contact. The larger particle size is found in the lubricant, the higher is it concentration in the contact. The work also shows the effect of changing the velocity on the change in concentration of the individual components and the connection of the transition velocity with the distribution of the components. The work gives a new perspective on the behavior of greases and provide useful knowledge for the development and optimization of greases for different applications.
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Membrane potential measurement with voltage sensitive dyes in fluorescence microscopy
Tkáč, Jan ; Janoušek, Oto (referee) ; Čmiel, Vratislav (advisor)
The aim of this work is to make a literature search in the measurement of membrane voltage using voltage-sensitive dyes and suggest a method for measuring the membrane voltage on the available cells using the voltage-sensitive dye di 4 ANEPPS and its further implementation. The work contains an introduction to electrophysiology of cells, and explains typical fluorescence characteristics. The thesis contains the description of a fluorescence microscope. The document presents characteristics of voltage-sensitive dyes and their distribution. A large part of the work describes the implementation and measurement of the experiment. The document also includes different methods for measuring and processing of all results.
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Cell Viability Measurement
Pelc, Pavel ; Janoušek, Oto (referee) ; Čmiel, Vratislav (advisor)
This work discusses the viability of cells. It describes the principle of fluorescence, fluorescent methods, fluorescent microskopy and the method of dyeing of the cells with a special kit that contains calcein AM and ethidium homodimer dyes. Further it describes the program for analysis and evaluation of viability on two different cellular cultures.
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Anisotropy techniques in study of cytoplasm
Sýkorová, Kateřina ; Obruča, Stanislav (referee) ; Mravec, Filip (advisor)
The main goal of this thesis was to compare experiments using time-resolved anisotropy and steady-state anisotropy for measuring in bacteria strain Cupriavidus necator. Fluorescent probe for anisotropy imaging was chosen BCECF_AM, which is derivate of fluorescein. Using experiment in system glycerol/water with fluorescein, anisotropy has been verified and calculated molecular hydrodynamic volume of a single fluorescein molecule, which approximately corresponded with real value. By using fluorescence imaging anisotropy microscopy, images and values of average anisotropy in cells were taken. Images of living cells (bacteria) of CN H16 and mutant CN PHB-4 showed differences, mainly in the uniformity of the inside environment.
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Natural drugs in cancer treatment and their cytotoxicity
Hájková, Tereza ; Raudenská, Martina (referee) ; Provazník, Ivo (advisor)
The thesis deals with the natural substances in context with the cancer disease. The natural substances have a positive effect on the human organism and they are able to influence the viability and the growth of the cancer cells. The main mechanical device is to influence the mechanisms needed to start the apoptosis of the cancer cells and stopping further proliferation. The cancer cell lines utilization in the cancer disease is discussed in the thesis too. The thesis states common methods of determining the natural substances cytotoxicity. For the experimental part of the thesis it was chosen the MTT test method and the xCELLigence system for monitoring in real time. The mechanical device of the tested substance capsaicin in application on the prostate cell lines, tumorous PC3 and nontumorous PNT1A influence will be observed within the experimental part of the thesis.
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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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Membrane potential measurement with voltage sensitive dyes in confocal microscopy
Heczková, Monika ; Sekora, Jiří (referee) ; Čmiel, Vratislav (advisor)
The aim of this work is to make a literature search in the measurement of membrane voltage using the voltage-sensitive dyes and suggest a method of measuring the membrane voltage on the available cells using the voltage-sensitive dye Di-4-ANEPPS and RH237. The work contains an introduction to electrophysiology of cell, explains fluorescence and typical fluorescence characteristics. The thesis contains the description of a fluorescence microscope. The document was largely devoted to characterization and distribution of voltage-sensitive dyes. The output of a design solution is a real experiment.
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Fluorescence imaging techniques in multimodal holographic microscope
Vašíček, David ; Procházková, Jana (referee) ; Čolláková, Jana (advisor)
The diploma thesis deals with the registration of images taken with the multimodal holographic microscope (MHM). The summary covers the fluorescent and holographic microscopy, and the multimodal holographic microscope combining both these microscopy types. Every pair of the images needs to be aligned in order to gain new information by combining both image types. The thesis contains an algorithm that registers images by phase correlation as well as a process created in MATLAB in accordance with the algorithm. The most important procedure parameters’ influence on the registration success is described and the results are annotated.
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