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Use of advanced fluorescence methods in the study of aggregation
Uhrínová, Alžbeta ; Enev, Vojtěch (oponent) ; Venerová, Tereza (vedoucí práce)
The diploma thesis deals with the use of advanced fluorescence techniques in the study of aggregation of selected aggregation models (casein and sodium dodecyl sulfate). At the beginning of the research, fluorescent probes Atto 488 and Rhodamine 6G (RH6G) were characterized using a spectrofluorometry, UV-VIS spectrometry, time-resolved fluorescence, and fluorescence correlation spectroscopy (FCS). Aggregation models with the addition of fluorescent probes were characterized using the same advanced fluorescence techniques. The interaction between the fluorescent probe Atto 488 and the aggregation models was not confirmed. The excitation maximum of Atto 488 was at 500 nm and the emission maximum at 520 nm. The lifetime of the fluorescent probe Atto 488 was from 4,20 to 4,82 ns and the hydrodynamic radius was determined to 0,6 nm. Rhodamine 6G had an excitation maximum at 525 nm and an emission maximum at 555 nm. The lifetime of the fluorescent probe RH6G was from 3,99 to 5,27 ns. The intensity of RH6G was found to decrease upon addition of a higher concentration of casein, resulting in quenching. The hydrodynamic radius of the RH6G probe was determined to 0,6 nm. With the addition of casein using the maximum entropy method (MEM) evaluation was from 0,5 to 534 nm. Stable SDS micelles had a hydrodynamic radius by MEM evaluation of 2,4 nm.
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Isolation of photosystem II particles and their characterization
Svitačová, Denisa ; Pekař, Miloslav (oponent) ; Venerová, Tereza (vedoucí práce)
This master's thesis deals with the isolation and characterization of spinach thylakoid membranes and photosystem II particles. Based on the research, two procedures were chosen for the isolation of spinach thylakoid membranes and photosystem II particles with an emphasis on preserving their native properties. Subsequently, the characterizations were carried out, focusing on the spectroscopic properties of the isolated samples. UV-VIS spectrometry was used to determine the ratio of chlorophyll a to chlorophyll b in samples of thylakoid membranes and photosystem II particles. The success of the isolations was confirmed by fluorescence methods. The excitation and emission spectra of the samples were measured by the steady-state fluorescence spectroscopy, and the fluorescence lifetimes of the samples of thylakoid membranes and photosystem II particles were determined by time-resolved fluorescence. The isolated samples were afterwards visualized with a fluorescence microscope with a CCD camera and the FCS technique.
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Influence of plasma treated water on the plant growth and vitality
Vozár, Tomáš ; Zahoranová, Anna (oponent) ; Kozáková, Zdenka (vedoucí práce)
This diploma thesis deals with the effect of plasma activated water on plant growth and vitality. By interaction of plasma with water, water acquires new unique properties that can be used in agriculture. The theoretical part deals with the basic properties of plasma, possibilities of preparation of plasma activated water in laboratory conditions, its physico-chemical characterization, and possibilities of its use in agriculture. In the next part, plant photosynthesis is discussed in connection with chlorophyll fluorescence, which measures the activity of photosystem II. Activity is an indicator of plant vitality, under environmental changes. At the end of the theoretical part, there are listed further possibilities for the analysis of plant material and food, such as elemental analysis, sensory analysis, and determination of dry matter content. Within the experimental part, plasma activated water (PAW) was prepared using plasma nozzle with introduced gas mixture that was immersed in the treated liquid. Further, plasma activated water was characterized and applied to radish plants. The effect of PAW on the plant growth and vitality was observed. The effect on the growth was observed through determination of fresh weight and dry matter content, the effect on vitality was assessed through chlorophyll fluorescence and the NDVI index. In the end, the effect on the content of stored elements C, H, O, and N in plants was determined and a sensory analysis was performed. The results show positive effects of plasma activated water on the plant growth and its sensory parameters such as appearance, taste, or spiciness. The results further show that PAW does not adversely affect plant vitality. This experiment should be continued by further indoor replication of the experiment to confirm obtained results and then, by moving the experiment to exterior in a form of field experiments.
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Pharmacokinetics of Intramuscularly Administered Thermoresponsive Polyacrylamides
Groborz, Ondřej
5 Farmakokinetika intramuskulárně aplikovaných termoresponzivních polyakrylamidů Autor: Ondřej Groborz Vedoucí práce: doc. Mgr. Martin Hrubý, Ph.D., DSc. Ústav makromolekulární chemie, Akademie věd České republiky Konzultanti: Ing. Pavel Švec RNDr. Lenka Loukotová, PhD. Abstrakt Roztoky některých polymerů vykazují dolní kritickou rozpouštěcí teplotu (lower critical solution temperature, LCST). Takové polymery za určitých podmínek vytvářejí homogenní roztoky, ale jakmile teplota roztoku překročí kritickou teplotu (cloud point temperature, TCP), dojde k fázové separaci. Pro tyto polymery bylo navrženo nespočet aplikací v rámci medicinální chemie a medicíny, například in situ formovaná depa pro radioterapii (brachytherapii), in situ formovaná depa pro protrahované uvolňování léčiv, imunoradiotherapie, injekční thermogelling pro tkáňová inženýrství a pěstování buněčných kultur, tracery pro magnetickou rezonanci (MRI) a mnoho dalších. Navzdory četným navrženým využitím a rozsáhlému výzkumu těchto polymerů, farmakokinetika injekovaných termoresponzivních polymerů stále není dostatečně dobře prozkoumána. V této práci se věnujeme syntéze a důkladné fyzikálně-chemické charakterizaci čtyř různých termoresponzivních polyakrylamidů, konkrétně poly(N-(2,2-difluoroethyl)akrylamidu, poly(N- isopropylakrylamidu),...
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Fluorescent Methods in Research of Eukaryotic Cells
Chmelíková, Larisa ; Babula, Petr (oponent) ; Pešl,, Martin (oponent) ; Provazník, Ivo (vedoucí práce)
This work examines the application of the fluorescent methods in use in in vitro studies to the field of cardiac tissue regeneration. Confocal fluorescence microscopy is an appropriate microscopic technique for studies in this field because it enables the visualisation of 3D structures and cell distribution in 3D models. The applied fluorescent markers should remain stable for a long period, are biocompatible, and are non-toxic for living cells. Nanoparticles such as superparamagnetic iron oxide nanoparticles (SPION) are currently very popular, and many studies have shown that they are suitable for long-term experiments. This research makes use of rhodamine-derived superparamagnetic maghemite nanoparticles (SAMN-R) and describes their excitation and emission spectra, size, and location within cells. A toxicity assay was performed by measuring reactive oxygen species (ROS) and non-quantitative measurements were conducted using fluorescence microscopy, confirming that a dose value of 20 µg·cm-2 is optimal for the treatment of living cells. This research also looks at the effects of SAMN-R treatment on cell proliferation and motility. The 3T3 fibroblast cell line was used for the cell proliferation test and scratch assay, after which human adipose-derived mesenchymal stem cells (MSCs) were used to examine single-cell migration. The subsequent statistical analysis revealed that it cannot be confirmed that the SAMN-R treatment exerts a significant effect on either cell proliferation or collective and single-cell migration, and it can be assumed that SAMN-R are appropriate fluorescent cell markers for living cells research, including the field of regeneration studies. Adipose-derived MSCs have enormous potential for cardiac tissue regeneration. Their interactions with HL-1 cardiac muscle cell line were studied using the scratch assay, and this model seems to be a promising and useful way to study cell-to-cell contact and its role in cell repairing.
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Metody vizualizace proudění
Ďurdina, Lukáš ; Jedelský, Jan (oponent) ; Lízal, František (vedoucí práce)
ĎURDINA, L. Metody vizualizace proudění. Brno: Vysoké učení technické v Brně, Fakulta strojního inženýrství, 2010. 45 s. Vedoucí bakalářské práce Ing. František Lízal. Práca pojednáva o metódach vizualizácie prúdenia tekutín s ohľadom na možnosti ich praktickej aplikácie. Chronologický opis historicky významných vizualizačných experimentov zdôrazňuje úlohu vizualizácie prúdenia v experimentálnej mechanike tekutín. Stručná charakteristika prúdového poľa obsahuje matematický popis objektov, ktoré môžeme v prúdení zviditeľniť. Vizualizačné metódy sú rozdelené na experimentálne a počítačovo asistované. Jednotlivé metódy sú radené postupne od tých s najnižšími nárokmi na vybavenie po náročnejšie metódy. Postupne sú v práci objasnené rôzne techniky, pomocou ktorých je možné získať kvalitatívne aj kvantitatívne údaje o sledovanom prúdení plynov a kvapalín. Poznatky sú následne zhrnuté do stručného prehľadu pre voľbu metód pre rôzne aplikácie. Na záver je uvedený popis vykonaného jednoduchého experimentu vizualizácie prúdenia pomocou metódy héliových bublín.
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Animovaný grafický model
Mikšaník, Martin ; Kučiš, Michal (oponent) ; Zemčík, Pavel (vedoucí práce)
Bakalářská práce se zabývá demonstrací málo používaných optických jevů v počítačové grafice, konkrétně fluorescencí a dispersí světla. Práce také navrhuje jiný postup při práci se světlem, materiálem a jeho vykreslení. Výsledný návrh je realizován v přiloženém inter- aktivním demonstračním programu.
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Měření membránového napětí pomocí napěťově citlivých barviv ve fluorescenční mikroskopii
Tkáč, Jan ; Janoušek, Oto (oponent) ; Čmiel, Vratislav (vedoucí práce)
Cílem této práce je sestavit literární rešerši v oblasti měření membránového napětí pomocí napěťově citlivých barviv a navrhnout postup měření membránového napětí na dostupných buňkách s využitím napěťově citlivého barviva di-4-ANEPPS a realizovat ho. Práce obsahuje úvod do elektrofyziologie živočišných buněk, vysvětluje fluorescenci a její typické vlastnosti a věnuje se popisu fluorescenčního mikroskopu. Dokument se z části věnuje charakterizaci a rozdělení napěťově citlivých barviv. Velká část práce obsahuje popis realizace a měření experimentu. V dokumentu jsou také zahrnuty různé metody měření a zpracování všech výsledků.
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Měření viability buněk
Pelc, Pavel ; Janoušek, Oto (oponent) ; Čmiel, Vratislav (vedoucí práce)
Tato práce se zabývá viabilitou buněk. Popisuje princip fluorescence, fluorescenční metody, fluorescenční mikroskopii a metodu barvení buněk pomocí speciálního kitu obsahující barviva calcein AM a ethidium homodimer. Dále je zde popsán program pro analýzu a vyhodnocení viability na dvou odlišných buněčných kulturách.
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Defect detection on fiber materials using machine learning
Lang, Matěj ; Richter, Miloslav (oponent) ; Honec, Peter (vedoucí práce)
Following Master's Thesis is presenting the creation of scanning unit, that will automate the quality check process in company SILON, for their fibered material. The process of manual detection is discussed and the automated solution is proposed. Several test are shown, that demonstrate effects of different lights on fibres dyed in Rhodamine. Optimal filter for camera is chosen, to achieve images with highest resolution possible and with enough definition. Next, the software tools for hardware control are presented and tools for building neural networks. Also, some basic info on current state of the art is provided, to explain some of the tools used. The network itself is shown and also its learning process and capabilities of defect detection.
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