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Optimization of surface-enhanced Raman scattering spectroscopy for study of biologically important biomolecules and their interactions
Šmídová, Natália ; Procházka, Marek (advisor) ; Vlčková, Blanka (referee) ; Michl, Martin (referee)
Title: Optimization of surface-enhanced Raman scattering spectroscopy for study of biologically important biomolecules and their interactions Author: Natália Šmídová Department: Institute of Physics of Charles University Supervisor of the doctoral thesis: Doc. RNDr. Marek Procházka, PhD. Abstract: The main goal of this thesis was to optimize surface-enhanced Raman scattering (SERS) spectroscopy for study of biologically important biomolecules. For that purpose we focused on substrates based on gold colloidal nanoparticles immobilized to silanized glass plates. Stable, uniform and highly reproducible SERS-active substrates have been prepared by using aminopropyltrimethoxysilane and citrate- reduced gold nanoparticles thermally stabilized after their immobilization. Model biomolecules 5,10,15,20-tetrakis(1-methyl-4-pyridyl)porphyrin (TMPyP) and 5,10,15,20-tetrakis(4-sulfonatophenyl)porphyrin (TSPP) were studied on these substrates by using a classical Raman spectrometer in macro-mode and a confocal Raman microspectrometer. Conditions for SERS spectroscopy of porphyrins were optimized with respect to sensitivity and reproducibility. SERS microspectroscopy showed several advantages over SERS measurements in macromode: possibility of surface spectral mapping, easier manipulation with samples, shorter...
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Development and optimization of systems for SERS on single molecule level
Michlová, Magdalena ; Vlčková, Blanka (advisor) ; Pfleger, Jiří (referee) ; Procházka, Marek (referee)
AABBSSTTRRAACCTT Dimers and small aggregates as well as compact aggregates of Ag nanoparticles (NPs) were assembled and chemically anchored to supporting surfaces. The supporting surfaces were either glass slides or SiO2 - coated Cu or Au grids for TEM, both chemically functionalized by 3-aminopropyltrimethoxysilane (APTMS). Compact aggregates of Ag NPs incorporating protoporphyrin IX (PPIX) molecules were prepared by adsorption of chlorides in the presence of PPIX. Dimers and small aggregates of Ag NPs were assembled by selected molecular linkers: 4,4'-diaminoazobenzene (DAAB), 4,4'-diaminoterphenyl (DATP) and 5,10,15,20-tetrakis(4-aminophenyl)porphine (TAPP). The most efficient strategy of dimers and small aggregates preparation has been their assembling by a three - step procedure involving (i) attachment of isolated Ag NPs to the NH2 groups of APTMS functionalized TEM grid, (ii) attachment of molecular linker (with two functional NH2 groups in para position) to Ag NPs by a one terminal NH2 group, and (iii) attachment of Ag NPs to the second, free terminal NH2 group of the linker. In this procedure, the control over the perpendicular orientation of the bifunctional linker and its attachment by one terminal group to Ag NP surface has been accomplished by functionalization of Ag NPs by adsorbed citrate...
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Application of Raman spectroscopy for study of nitrogen containing compounds for astrobiological purposes
Culka, Adam ; Jehlička, Jan (advisor) ; Vlčková, Blanka (referee) ; Frank, Otakar (referee)
IV ABSTRACT This PhD thesis was focused on the evaluation of the application of Raman spectroscopy as an analytical method used specifically for research on selected nitrogen containing compounds in experiments, that are important in the astrobiological context. The results of experiments provided insight into the advantages as well as the limitations of the method in several applications that are expected to be encountered in the future planetary exploration missions with astrobiological context use, and where the Raman spectroscopy is proposed as an advantageous method. The Raman spectroscopy was tested in various experimental tasks. Several nitrogen containing minerals were analysed, where the features such as the indestructive analysis, small laser spot size, and selection of the excitation source enabled to acquire spectra of minerals even with miniature sample sizes, also the band assignment was published for the first time for some of the studied minerals. Artificially prepared samples of biomarkers mixed in mineral matrices were analysed to test the ability of the method to analyse complex mixtures: three biomarkers and two evaporitic minerals were analysed with a portable instrument, and all compounds were unambiguously detected. When the samples contained more of the similar compounds with many...
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Surface-enhanced resonance Raman scattering of Zn(II) porphyrins in systems with aggregated and non-aggregated Ag nanoparticles
Spáčil, Dušan ; Vlčková, Blanka (advisor) ; Procházka, Marek (referee)
In this diploma thesis, SERRS(Surface - enhanced resonance Raman Scattering) and SERS (Surface - enhanced Raman Scattering), surface of plasmon extinction spectra and TEM images of systems with tetracationic zinc porphyrin ZnTMPyP and silver nanoparticles (Ag NPs) were studied and interpreted. The systems with isolated nanoparticles and the systems with compact aggregates were investigated. The systems with isolated nanoparticles were prepared by addition of ZnTMPyP to hydrosol of Ag NPs. NaCl was added to this system and so compact aggregates were prepared. SERRS and SERS spectra of ZnTMPyP were studied at excitation wavelengths λexc= 441.6 nm a 532 nm. Firstly, stationary systems and secondly dynamic development of hydrosol Ag NPs → Ag NPs/ ZnTMPyP → Ag NPs/ ZnTMPyP/ NaCl systems for 680 s was investigated. Time evolution of SERRS and SERS spectra were analyzed by factor analysis (FA) and time evolution of spectra extinction plasmon were measured. FA showed a strong increase of the signal of ZnTMPyP after the addition of NaCl, i.e. after conversion of isolated nanoparticles to compact aggregates. In these systems with high concentration of ZnTMPyP the increase of signal was succeeded by its decreased. With help of FA the limit of SERRS spectral detection (λexc= 441,6 nm) and limit of SERS spectral...
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Bipyridine complexes of Ru(II) as SERS spectral probes of the mechanisms of surface -enhanced optical processes
Kokošková, Markéta ; Vlčková, Blanka (advisor) ; Šmejkal, Petr (referee)
SERS and SERRS (surface enhanced /resonance/ Raman scattering) spectra of a dicationic Ru (II) bis(2,2'-bipyridine)(4,4'-dicarboxy-2,2'-bipyridine) /Ru(bpy)2(dcbpy)/ complex in systems with aggregates of unmodified and/or chloride-modified Ag nanoparticles (NPs) were obtained and compared to those of dicationic Ru (II) tris(2,2'-bipyridine) /Ru(bpy)3 /. The splitting of several bands and the appearance of a new band at 1367 cm-1 observed solely in SERS and SERRS of Ru(bpy)2(dcbpy) in the system with unmodified Ag NP aggregates was attributed to chemisorption of the complex onto Ag NP surface via two carboxylate groups. SERS/SERRS excitation profiles obtained for the spectral bands and attributed to the Ru- dcbpy unit of the chemisorbed Ru(bpy)2(dcbpy)/ complex were found to maximize at 488 nm excitation, while those of the two Ru-bpy units peaked at 458 nm. Comparison of the profiles with the electronic absorption spectrum of free Ru(bpy)2(dcbpy) has revealed that chemisorption of the complex causes a red-shift of the Ru→dcbpy charge transfer transition band. The observed decrease of the energy of the Ru→dcbpy charge transfer is explained by an increase of the electron-withdrawing ability of the two COO- groups upon their chemisorption on AgNP surface. Concentration value of SERRS spectral...
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Graphical support for entering a Unix command line
Podzimek, Andrej ; Vlčková, Blanka (referee) ; Rindt, Eduard (advisor)
The present work comprises a detailed description and documentation of a software project called GRUX. The main purpose of the software is to provide an interactive alternative to man pages. Its input data structure describes a hierarchy of widgets and relations among them. The program outputs code written in the Java programming language, using the SWT (Standard Widget Toolkit) library to display a graphic dialogue window. This work gives a detailed description of input and output data structures and the implementation of the dialogue window generator. The work also includes all the documentation needed for both control and further development of the software.
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Guesthouse
Vlčková, Bára ; Kučera, Radim (referee) ; Pěnčík, Jan (advisor)
This bachelor thesis is a project of new five-storey, semi basement guesthouse in Opava. This guesthouse will be used for accommodation, catering and recreation. Accommodation capacity is 21 people. In the basement is a technical background. The first floor is designed for the overall run of the guesthouse, staff, catering and sports. In the second and third floor are rooms for guests. Almost every room has a balcony. In the fourth floor is a common room with adjoining terrace. The building is brick. The vertical structures are made of ceramic blocks. Outer walls are insulated by outer contact system. The ceilings are mostly ceramic-concrete and a smaller part of the reinforced concrete. The guesthouse is roofed by single-flat roof.
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Testing of commercial products for separators of lipids
Vlčková, Bohumila ; Vítová, Eva (referee) ; Omelková, Jiřina (advisor)
Diploma thesis was aimed at testing of five commercial products for separators of lipids. Performed study should to shown that in produts are presented lipase-produducing microoganisms, event. that in products are also lipolytics enzymes as addition. The products was cultivated by solid-state and submerged fermentations and subsequently was tested their properties as is lipolytic activity, the amount of free fatty acids etc. Solid-state fermentation used Tributyrin Agar Base where Tributyrin was used as a substrate. Submerged fermentation used special prepared medium where was olive oil. Methods using in this study served for detection and qualification of lipase-produducing microoganisms in individual products. For detection of lipase-produducing bacterial spieces was used Spirit Blue Agar. In this case was used Tributyrin and Tween 80 as a substrate. Lipolytic activity was measured by spectrophotometry and degradation of lipids was measured titrimetry. Performed study demonstrated that all tested products have ability to degradation of lipids.
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FAMILY HOUSE IN ŠTÍTINA
Vlčková, Bára ; Dohnal, Jakub (referee) ; Pěnčík, Jan (advisor)
This bachelor thesis is project of two-storey cellar family house in Štítína. The house is bricked and it’s designed for four-member family. In the basement is common room, food storage, dryer room, toilet, technical room and bike room. On the first floor is vestibule, living room with kitchen, bathroom, toilet and guest room. Garage for one car is connected to the object. Garage it’s not operationally connected with living part. On the second floor we find two children rooms, bedroom, bathroom, toilet and workroom. One children room has balcony access. Second floor is roofed by gabled roof. Above the garage is flat roof. Project is processed with computer program AutoCAD.
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Art commerce in Czech Republic
Vlčková, Barbora ; Drozen, František (advisor) ; Novák, Miloš (referee)
Bachelor thesis describes art commerce in Czech Republic with emphasis on paintings. It compares this situation to the situation on foreign markets and proposes how to improve it. This thesis also describes how auction houses and galleries work, laws concerning the art commerce and support of art in Czech Republic.
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