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Complementary analysis of procaryotic cells by electron microscopy and Raman spectroscopy
Ikrényiová, Terézia ; Sedláček, Petr (referee) ; Hrubanová, Kamila (advisor)
This master thesis deals with conventional methods of bacterial cell analysis, polyhydroxyalkanoates, Raman spectroscopy and electron microscopy in the theoretical part. The production of polyhydroxybutyrate by selected thermophilic bacteria and their analysis by gas chromatography, cryogenic scanning electron microscopy and Raman spectroscopy is described in the experimental part. The chosen sample was analyzed by a transmission electron microscope. Comparing the results from previous mentioned methods it was found that the bacteria Schlegelella thermodepolymerans accumulated the highest amount of PHB. The lowest amount of PHB was obtained by bacteria Rubrobacter xylanophilus. The assumption that the PHB granules formed so-called needle-like plastic deformations during freeze-fracturing was affirmed by cryo-SEM photos analysis. Moreover, it was found that the bacterial cell characterization deduced from microscopic observation of samples corresponded to the description in the literature. TEM provided better resolution photos and in consequence the cells and PHB are more visible. The thesis is also focused on chemical fingerprint analysis of cells by Raman spectroscopy. Several biomolecules were identified by measured Raman spectra for the particular samples.
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Optical micromanipulation and Raman spectroscopy of cells in microfluidic systems
Klementová, Tereza ; Samek, Ota (referee) ; Mravec, Filip (advisor)
This diploma thesis deals with optimization of analysis process and measuring antibiotics induced changes in E. coli cells via Raman spectroscopy, LTRS and microfluidic systems. Optical micromanipulation by a laser beam allows noncontact and noninvasive manipulation of objects on scale 10^-5–10^-8 m, for example bacterial cells. Microfluidic device consists of microchannels and microchambers in transparent polymer and it is used for isolation, observation and cultivation of bacterial cells. Combination of these methods gives an effective tool for observation, manipulation and analysis of microorganisms. E. coli is a microorganism potentially pathogenic for humans and faster detection of its sensitivity to antibiotic treatment would make the whole process of diagnostics and treatment easier. We performed laser tweezer-Raman spectroscopy and conventional Raman spectroscopy of bacterial cells and cells under antibiotic stress and collected Raman spectra and characteristic areas were compared with literature to establish the reliability and usefulness of this method.
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Selective hydrogenation/Oxidation of CVD graphene using AFM
Přikryl, Vojtěch ; Švarc, Vojtěch (referee) ; Bartošík, Miroslav (advisor)
Graphene is a perspective material for utilization in electronics due to its high charge carrier mobility. However, in order to be applicable in practice the bandgap opening is necessary. One way of achieving this is modification of graphene by hydrogenation or oxidation. This work concerns the possibility of using AFM for local hydrogenation/oxidation of graphene depending on the bias voltage between the tip and the sample. The presence of hydrogenation/oxidation was examined by Kelvin probe force microscopy and Raman spectroscopy. The possibility of oxidation was proved but hydrogenation was not proved.
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Study of the morphology of highly oriented pyrolytic graphite irradiated by argon ions
Komínek, Josef ; Kaspar, Pavel (referee) ; Sobola, Dinara (advisor)
The main task of this bachelor’s degree work is study of highly oriented pyrolytic graphite (HOPG) using atomif force microscopy, Raman spectroscopy and XPS spectroscopy. HOPG is layered material: atoms in one layer have stronger bong between themselves than two atoms in two neighbouring layers. Wide application options require further study. HOPG is now commonly used in nano-industry. It is used as a dimension standard. HOPG of A quality was processed in this work. In order to get accurate 3D data of sample surface, AFM measuring was done. 3D models of surfaces of various sizes are made. For these purposes was used free AFM data tool. Height parameters are used to describe topography. Got parameters are important for further study.
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Raman spectroscopy teaching equipment
Indra, Zdeněk ; Sedlák, Petr (referee) ; Škarvada, Pavel (advisor)
This thesis examines the principle of the Raman phenomenon and the function of the Raman spectrometer in order to build a low-cost Raman spectrometer assembly for school laboratory teaching. The thesis lists the important parts of the device and describes their key features. Subsequently, the selection of each component is discussed and a basic experiment is performed to verify the principle of this spectrometer. In the final part, a low-cost Raman spectrometer is constructed and tested thanks to the knowledge gained.
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Raman spectroscopy of biologically active species and antitumor drugs
Třeštíková, Liběna ; Vetterl,, Vladimír (referee) ; Vrána, Oldřich (advisor)
SERRS spektra of biological materials are very komplex, because they consist of signals from all molecules present in cells. In this text are presented SERRS spektra of antitumor drugs and its komplex with DNA. Experimental are rated on doxorubicin and another antitumor druha and on study of theirs potential by treatment for tumors. Doxorubicin belong to clase antracycline antibiotics and is used for stop of tumor cells reproduction. Scientists found still new ways, new drugs. SERRS is one of possibilities for study of this drugs and theirs interaction with DNA.
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Molecular modelling - Structure and Properties of carbene-based catalyst
Kulovaná, Eva ; Hermanová, Soňa (referee) ; Richtera, Lukáš (advisor)
Pomocí molekulového modelování je možné předpovídat chování nových látek a napomáhá při jinak obtížné interpretaci experimentálních dat. Cílem práce byla predikce vybraných vlastností polymeračních katalyzátorů na bázi karbenů, predikce jejich struktur a spektrálních charakteristik a studie mechanismu polymerace za otevření kruhu laktidu. K ověření chování karbenů a jejich prekurzorů ve formě chloridů byly studovány vybrané charakteristiky molekuly. Byl proveden výpočet vybraných molekulových orbitalů a elektrostatických map. Následně pomocí počítačových programů byly získány teoretické vazebné délky a úhly vybraných imidazolových a imidazolinových sloučenin, karbenů a jejich možných produktů hydrolýzy. Data strukturně podobných, již charakterizovaných sloučenin, byla získána z CCDC (Cambridge Crystallographic Data Centre) a následně byla konfrontována s vypočítanými daty. Byla změřena infračervená a Ramanova spektra imidazolové soli a infračervené spektrum příslušného karbenu. Tato spektra byla konfrontována s napredikovanými. Pro lepší interpretaci spekter byla spočítána spektra možných produktů hydrolýzy. Následně byl studován mechanismus polymerace za otevření kruhu laktidu. Na základě spočítaných energií stacionárních bodů byl navržen nový mechanismus polymerace.
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Preparation of graphene layers by various methods and characterization of their properties
Zahradníček, Radim ; Bartošík, Miroslav (referee) ; Lišková, Zuzana (advisor)
This bachelor's thesis deals with the production of graphene by different methods. Graphene was prepared by mechanical exfoliation and chemical methods. Graphene oxide (GO) was first prepared by modified Hummers method and disolved the solution in isopropyl alcohol. It was subsequently reduced by photoreduction, UV radiation and thermal reduction. Produced graphene samples were characterized using optical microscopy, electron microscopy, atomic force microscopy, Raman spectroscopy and X$-$ray photoelectron spectroscopy. Measurement results and evaluation of production quality of graphene are shown in this work.
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Characterization of graphene electrical properties on MEMS structures
Brodský, Jan ; Pekárek, Jan (referee) ; Gablech, Imrich (advisor)
This work presents basic properties of graphene, methods for its synthesis and methods for its characterization by Raman spectroscopy, two-point probe, four-point probe and van der Pauw measurements. The experimental part of this work describes the process of graphene oxide, reduced graphene oxide and graphene sample preparation and measurement of its current-voltage characteristic by two-point probe method. Subsequently, sample annealing in vacuum furnace is described in this work. The annealing is important for acquiring good electrical contact between the 2D material and electrodes. Sample analysis by Raman spectroscopy is performed. The last chapters of this work describe design and fabrication of MEMS structure. Such structure serves for characterization of graphene and other 2D materials.
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Raman spectroscopy as a tool for analysis of biotechnologically relevant microorganisms
Záhorská, Linda ; Enev, Vojtěch (referee) ; Mgr.Ota Samek, Ph.D. (advisor)
The diploma thesis deals with the study of biotechnologically significant microorganisms, using the Raman spectroscopy. Content of the theoretical part is brief characteristic of Raman spectroscopy as a method, its use in practice and also use as a tool for monitoring of biotechnologically processes. Thesis was further focus on the biotechnologically significant microorganism Aureobasidium pullulans, its use in biotechnology and also for over-produced substances and in particular poly-L-maleic acid and pullulan. The content of the experimental part was study of selected strains A. pullulans, specifically stains as DSMZ, CCM F148 and CCM 8182, using Raman spectroscopy on the various types of culture media. Subject of practical part research was too production of extracellular polymers, acid poly-L-apple and pullulan, by selected strains A. pullulans. Objective of my thesis was described and determinate, spectra of individual strains as well as extracellular products, mainly pullulan, and then choose suitable production medium and optimal production strain A. pullulans. During experimental work was found, that optimal production strain was DSMZ strain culture on the mineral medium with the addition of yeast autolysate, which was optimal medium type. The content of the pullulan produced was for gravimetric determination, 6,3g/L, which also confirmed the results of the HPLC method. It was experimentally found, that Raman spectroscopy isn´t suitable method for quantification of extracellular products, but is appropriate and was used for PCA analysis of individual strains.
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