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Robot Localization Using Camera
Heřman, Petr ; Španěl, Michal (referee) ; Beran, Vítězslav (advisor)
The objective of this work is to design a simple localization method and its implementation in robot operating system ROS. This method uses a monocular camera as the only sensor and estimates the position in a known map. In experiments with prototypes are tested key points of type SURF, SIFT and ORB.
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Robot Controlled by PIC Microprocessor Unit
Heřman, Petr ; Luža, Radim (referee) ; Rozman, Jaroslav (advisor)
This thesis describes design of a cheap robot. It includes implementation of firmware of low level control unit based on microcontroller PIC. The firmware drives motors, gains sensors data and communicates with the high level control unit. Furthermore the thesis presents realisation of connection to the robotic operation system ROS and its standard structures allowing usage of existing packages for the robot teleoperation and displaying sensor data on the remote computer. The thesis finally reports experiments with the robot. The constructed prototype is the model of the robotic lawn mower, however the whole solution has universal usage.
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Cellular protein interactions studied by advanced fluorescence imaging methods
Belejová, Sára ; Heřman, Petr (advisor) ; Krůšek, Jan (referee)
This thesis studies an important tumor suppressor, p53, and its interac�on partner, nucleophosmin (NPM), in living cells. Proteins are studied using fluorescence confocal microscopy techniques such as fluorescence life�me imaging and fluorescence anisotropy measurements. The primary focus of the research is on a specific variant of the p53 protein called p53-L344P, which is generated by a point muta�on from its original form (p53wt). We inves�gate the oligomeriza�on state of p53-L344P in vivo, which appears to be monomeric, confirming the results of in vitro experiments from other studies. Further, we show that p53wt and p53-L344P can form complexes with each other. We compare the interac�on of the NPMmutA protein with p53wt and p53-L344P proteins. Our findings reveal that the L344P mutant is not transferred from the nucleus to the cytoplasm in the presence of NPMmut, as is p53wt. Furthermore, we inves�gate the oligomeriza�on state of p53wt when it is in the cytoplasm and propose avenues for further research into this interac�on.
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Tracking membrane permeabilization on single lipid vesicles - method development.
Gücklhorn, David ; Šachl, Radek (advisor) ; Heřman, Petr (referee)
Protein complexes are challenging systems to study, especially when these complexes form on lipid membranes only for a short period of time. This is also the case of fibroblast growth factor 2 (FGF2), a protein that has many physiological and pathological functions in the human organism. It plays major role in the development of cancer as it promotes cell survival and angiogenesis. It also serves as a basis for development of novel treatments of nerve injuries. Despite being heavily studied for many years, it remains unclear how the protein is translocated into the extracellular space where it performs its function. To study complex systems such as FGF2 that self-assembles on the membrane into membrane penetrating pores we decided to develop a simple and efficient fluorescent microscopy method. This method is called double leakage single GUV assay (DLSGA). It utilizes giant unilamellar vesicles (GUVs) mimicking native cellular membranes. In a single experiment, up to 300 individual GUVs are imaged for the content of a leakage dye that reports on the presence of FGF2 pores. During three measurements and under different conditions, detailed information about pore-opening dynamics is gained for each GUV. Results of these measurements are then used to divide GUVs into six groups based on formation and...
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Properties and interactions of nucleoproteins studied by advanced lifetime-based imaging methods
Vacková, Anežka ; Heřman, Petr (advisor) ; Krůšek, Jan (referee)
The main aim of this thesis was implementation and testing of an additional imaging method for investigation of protein-protein interactions in our laboratory and to extend already established heteroFRET (Förster resonance energy transfer) method used with the fluorescence lifetime imaging (FLIM). We performed homoFRET experiments with both the steady-state and time-resolved anisotropy-imaging methods. The presence of homoFRET between fluorophores was done by partial fluorophore bleaching and by the "red edge" analysis. We compared the methods, especially their sensitivity, experimental requirements, complexity, and informational value. We effectively applied steady-state and time-resolved anisotropy imaging to six proteins of interest; both proved comparably effective and reliable as heteroFRET-FLIM for detection of the protein interactions. The red edge method appeared problematic due to background contribution, which could not be suppressed with the current setup and instrumentation. We validated steady-state and time-resolved anisotropy imaging by studying well- described oligomerizing nucleophosmin variants (NPMwt and NPMmut) and noninter- acting mutant NPMcut, expressed in living HEK-293T cells. Then, we also probed nu- cleolin (NCL) and free fluorescent tags NowGFP and mVenus, aiming to find...
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Fluorescence Spectroscopy: Advanced methods and their defined applications in protein science
Pospíšil, Petr ; Hof, Martin (advisor) ; Heřman, Petr (referee) ; Polívka, Tomáš (referee)
The hydration and dynamics of the biomolecules appear to be vital for their proper biological functioning. In the presented thesis, various fluorescence techniques were developed and applied to access these properties and their changes upon the mutual interactions of the biomolecules. Initially, the solvent relaxation method based on recording time-dependent fluorescence shift (TDFS) was used to map DNA interactions with proteins and lipids by the newly synthesised fluorene dye covalently bound to the DNA. Secondly, copper-transporting ATPase was probed by Badan attached to the copper-binding cysteine-proline-cysteine motif. The variations in hydration were found to be crucial for the proper ATPase function. Third, a detailed study on quenching of Badan/Prodan fluorescence by tryptophan revealed the limitations of the TDFS method for protein studies, which is essential finding for further applications of TDFS. Fourth application involves investigations of heavy atom effects on the excited state relaxation processes by up-conversion approach in iodinated metallocorroles, which are promising dyes for biological imaging. The obtained findings shall help in further tuning of the optical properties of the corroles desired for the variety of applications. Finally, fluorescence correlation spectroscopy...
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The History of the Sokol Movement in Lysé nad Labem
Herman, Petr ; Kašpar, Ladislav (advisor) ; Zachariášová, Jana (referee)
Sokolská organizace má za sebou bohatou minulost a je navždy zapsána do historie tohoto státu. Z jejích řad vyšlo mnoho obránců naší republiky, ať už jako legionáři nebo bojovníci za svobodu na bojištích druhé světové války. Sokol z nich vychoval lidi s vynikající morálkou i fyzickou kondicí. Lyský Sokol vznikl jen pár let po "průkopnickém" Sokole v Praze a hned byl velký zájem o členství v jednotě. Bohužel tragickou nehodou ve městě, která vyvolala krizi, byla činnost jednoty záhy na 2 desetiletí neúmyslně pozastavena. Naštěstí obyvatelé nezapomněli a Sokol znovu ožil a dále sílil. Tentokrát organizaci do cesty vstoupila válka, a tak činnost ochabla. Po válce se vzchopil a zažíval svá nej lepší léta. S druhou světovou válkou byla činnost Sokola opět zastavena a tentokrát ani po skončení neměl příliš prostoru k rozletu, komunistická diktatura ho též shledala nepřípustným. Přesto se Sokolové nevzdali a přes mnoho nepravostí působil v ilegalitě. Revoluční rok 1989 mu opět otevřel brány. Tak jak byl Sokol v rozkvětu za první republiky, si však dnes můžeme nechat zdát, ačkoliv dle mého názoru jsou jeho myšlenky stále aktuální. Bohužel dnešní společnost, jak se zdá, upřednostňuje zřejmě jiné cíle a hodnoty.
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Deconvolution fluorescence microscopy of yeast cells
Štec, Tomáš ; Plášek, Jaromír (advisor) ; Heřman, Petr (referee)
Title: Deconvolution fluorescence microscopy of yeast cells Author: Tomáš Štec Department: Institute of Physics of Charles University Supervisor: prof. RNDr. Jarmoír Plášek, CSc., Institute of Physics of Charles Uni- versity Abstract: Fluorescence microscopy presents an fast and cheap alternative to more advanced imaging methods like confocal and electron microscopy, even though it is subject to heavy image distortion. It is possible to recover most of the original distortion-free image using deconvolution in computer image processing. This al- lows reconstruction of 3D structure of studied objects. Deconvolution procedure of NIS Elements AR program undergoes an thorough inspection in this diploma the- sis. It is then applied on restoration of 3D structure of calcofluor stained cell wall of budding yeast Saccharomyces cerevisiae. Changes of the structure of the cell wall during cell ageing are being examined. Cell wall of aged cells shows increased surface roughness and even ruptures at the end of cell life. Keywords: fluorescence, microscopy, deconvolution, NIS Elements AR, calcofluor, yeast, cell wall, ageing
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