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SMV-2023-57: EM analysis of microorganisms II
Hrubanová, Kamila
The aim of the contract research was the expert analysis of microbiological samples with biotechnological significance with the intention of describing the ultrastructure of the cells, which depends on the cultivation conditions of the industrial process. The collaboration included cryogenic preparation of samples, followed by microbial microscopy using low-temperature electron microscopy (cryo-SEM), and selected samples were processed for transmission electron microscopy (TEM). Furthermore, spectroscopic measurements were performed using Raman spectroscopy.
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SMV-2023-56: EM analysis of microorganisms
Hrubanová, Kamila
The aim of the contract research was the expert analysis of microbiological samples with the intention of describing morphological changes dependent on culture conditions. The collaboration included cryogenic preparation of samples, followed by microbial microscopy using low-temperature electron microscopy (cryo-SEM), and selected samples were processed for transmission electron microscopy (TEM). Furthermore, spectroscopic measurements were performed using Raman spectroscopy.
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Raman spectroscopy as a tool for analysis of microbial cells
Pokorný, Petr ; Enev, Vojtěch (referee) ; Samek, Ota (advisor)
This bachelor thesis is focused on the use of Raman spectroscopy for analysis of ectoine and hydroxyectoine presence in bacterial strain Halomonas elongata. Theoretical part compile characteristics of extremophiles with closer look on halophilic organisms and one of their main osmolytes, ectoines. Following by description of Raman spectroscopy method and its uses. Experimental part deals with cultivation of bacteria on different combinations of substrate and salt concentration in order to reach optimal production of PHA and for achieving the highest possible yield of biomass to be analysed by Raman spektroskopy. As the best substrate for PHA production turned out to be glukose along with salt concentration 30 g/l NaCl, where percentage representation of PHB makes 30,5229 %. As an ideal substrate for the highest yield of biomass proved to be sacharose. Bacteria that grew on sacharose with three different salt concentrations, 40, 70 and 100 g/l NaCl was measured along with Halomonas salina and Halomonas organivorans for comparison. Raman spectroscopy unfortunately turned out to be inadequate for measurement of this type of sample, thus we were not able to measure ectoine of hydroxyectoine presence in cells.
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Analysis of biological significant substances
Maděránková, Denisa ; Rychtárik, Milan (referee) ; Roleček, Jiří (advisor)
Selected methods of Raman spectroscopy, like surface-enhanced Raman spectroscopy and single molecule Raman spectroscopy, are described in this diploma work. The basis of two methods for numerical modelling of optical properties of micro- and nanoparticles are prefaced. The methods are Discrete Dipole Approximation and Finite Difference Time Domain. Micro- and nanoparticles are used in surface enhanced Raman spectroscopy and other nanospectroscopic methods. Further, the main instrumentation needed for Raman spectroscopy is described. The first part of experimental section of this work is numerical modelling of photonic nanojet that occures behind dielectric microparticles. This phenomenon leads to a new technique of confocal microscopy with Raman spectra measuring. The second experimental section contains results of Raman spectra measurement with beta-carotene and surface-enhanced Raman spectra of beta-carotene in silver-sol solution.
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Characterization of PVDF material in nanoscale resolution
Pisarenko, Tatiana ; Dallaev, Rashid (referee) ; Sobola, Dinara (advisor)
Tato práce se zabývá charakterizací nanovláken na bázi polyvinylidenfluoridu. Zaměření práce je na piezoelektrické vlastnosti vlákna, které jsou studovány metodou piezoelektrické silové mikroskopie. Takto byly měřeny dva typy odlišných vzorků, které se lišily v parametrech výroby. Odlišnosti vláken v jejich fázovém složení byly také zkoumány za využití Ramanovy spektroskopie a infračervené spektroskopie s Fourierovou transformací. Chemická analýza povrchu a jeho stavu proběhla pomocí rentgenové fotoelektronové spektroskopie. Různé uspořádání nanovláken spolu s jejich průřezem bylo pozorováno rastrovacím elektronovým mikroskopem za využití fokusovaného iontového svazku. Rovněž byla zkoumána smáčivost a kontaktní úhel povrchu vzorků s demineralizovanou vodou. Bylo zjištěno, že vyšší rychlost otáček válce během procesu elektrostatického zvlákňování má velmi významný vliv na jejich uspořádání a tím i na parametry ovlivňující tvorbu piezoelektrického jevu a dalších materiálových vlastností.
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Raman spectroscopy of prokaryotic cells
Večeríková, Paula ; Bernatová,, Silvie (referee) ; Samek, Ota (advisor)
The presented bachelor‘s thesis deals with the Raman spectroscopy of bacterial cells. This method proves to be very suitable for monitoring the culture conditions of these bacterial strains because it is essentially non-destructive and fast compared to gas chromatography. Due to it‘s non-destructive nature, it is possible to further cultivate the measured cells. This analytical method can also be used to sort and select individual cells with increased ability to produce PHB. This would enable the possibility to select the generation of so-called PHB superproducers. Polyhydroxybutyrate (PHB) belongs to the group of polyhydroxyalkanoates (PHA), which serve as a substitute for plastics made in the petrochemical industry, whose consumption is increasing, especially during a pandemic. Current biodegradable substitutes are based on starch, which, in turn, depletes nutritional material for humans and animals. This bachelor thesis can serve as a basis outline for reducing the cost of PHB production, because Raman spectroscopy can be used in cell culture as a sensor of PHB content in response to a bacterial strain. In the experimental part of the work, the Raman spectra of selected bacteria were measured, where the result showed the greatest response to PHB in Chelatococcus shambunathi and the lowest in the thermophilic isolate BZ (Paenibacillus sp.). Quantitative information on the content of PHA in bacteria was obtained by gas chromatography, where the highest content was the bacteria- Chelatococcus shambunathi and the lowest Termobacillus composti. The subject of the second part was the separation of a mixture of PHA-producing and non-PHA-producing bacterial cultures. The result clearly proves that the mixture is separable based on the intensity of the selected sorting parameter.
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Preparation of ultra sharp tips for STM TERS applications
Šilhan, Lukáš ; Man, Ondřej (referee) ; Jan, Vít (advisor)
The goal of this bachelor thesis is preparation of Au tips by electrochemical etching. The theoretical part deals with comparison of microscopy techniques for materials surface characterization and Raman spectroscopy. The experimental part is focused on the settings of optimal etching parametres to gain appropriate geometry of fabricated tips. Function of tips is verified by evaluation of data gained by experimental measuring with this tips. Prepared tips are suitable for applications such as STM and TERS.
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Molecular modelling - Structure and Properties of carbene-based catalyst
Kulovaná, Eva ; Zmrzlý, Martin (referee) ; Richtera, Lukáš (advisor)
Molekulové modelování umožňuje předpovídat chování nových látek a napomáhá při jinak obtížné interpretaci experimentálních dat. Záměrem našeho studia byla predikce vybraných vlastností polymeračních katalyzátorů na bázi karbenů, predikce jejich struktur a spektrálních charakteristik. K ověření chování karbenů a jejich prekurzorů ve formě chloridů byly studovány vybrané charakteristiky molekuly. Byla provedena vizualizace vybraných molekulových orbitalů a map elektrostatických potenciálů a elektronových hustot. Následně pomocí počítačových programů byly získány teoretické vazebné délky a úhly vybraných imidazolových a imidazolinových sloučenin a z nich připravených karbenů. Data strukturně podobných, již charakterizovaných sloučenin, byla získána z CCDC (Cambridge Crystallographic Data Centre) a následně konfrontována s námi vypočítanými daty. Byla změřena infračervená (IČ) a Ramanova spektra imidazolové soli a IČ spektrum příslušného karbenu. Tato spektra byla konfrontována s napredikovanými.
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Electrospun fibers based on PVDF and nylon
Černohorský, Petr ; Sobola, Dinara (referee) ; Papež, Nikola (advisor)
Polymer nanofibers used for the construction of triboelectric nanogenerator (TENG) and piezoelectric nanogenerator (PENG) are new and promising technologies for energy recovery. Thanks to the generation of electrical energy based on mechanical movement (deformation), these fibers can find application in the field of self-powered electronic devices. In this work, three nanofibrous structures of materials were prepared by electrostatic spinning: pure polyvinylidene fluoride (PVDF), pure polyamide-6 (PA6) and their mixed combination PVDF / PA6. Non-destructive analyzes such as Raman spectroscopy, FTIR, XPS and electron microscopy were used to study the properties of nanofibers. Analyzes confirmed the positive effect of electrostatic spinning of polymers on the support of the formation of highly polar crystalline -phase in PVDF and , -phase in PA6. The structure arrangement of the nanofibrous material and their defects were observed by scanning electron microscopy (SEM). Furthermore, the contact angle of the wettability of the liquid on the surface was measured for the materials, and the permittivity was measured to monitor the dielectric properties. The described results make the mixed material PVDF / PA6 very promising for further research in the field of nanogenerators and functional textiles.
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