|
|
Use of fluorescence methods for the study of protein interactions
Johaníková, Klára ; Bezděková,, Jaroslava (referee) ; Pavelicová, Kristýna (advisor)
The diploma thesis "Use of fluorescence methods for the study of protein interactions" is focused on the use of fluorescence methods for the study of protein interactions using electromigration methods and Förster resonance energy transfer (FRET). The aim of this work was to create a bioconjugate of metallothionein (MT) protein with quantum dots (QDs) and commercial dyes. FRET was subsequently studied between these conjugates. QDs were synthesized under UV light and conjugation with MT was performed via a carbodiimide zerolengthcrooss-linker (EDC / sulfo-NHS), which serves to activate carboxyl groups and allows bioconjugation of the ligand by covalent bonding. Due to the high proportion of cysteines in MT, this protein has a very high affinity for metals. It is also involved in scavenging free radicals and there are studies that show that MT is overexpressed in cancer cells. Attention was also paid to the study of MT dimerization, which leads to an understanding of oxidative dimerization of MT and thus can contribute to understanding the formation of free radicals in the body and to deepen the knowledge about neurodegenerative disorders such as Parkinson's or Alzheimer's disease or amyotrophic lateral sclerosis. The formation of the MT dimer was confirmed by energy transfer between the donor (QDs) and the acceptor (commercial dye-cyanine) through the physical phenomenon of FRET and MALDI-TOF-MS.
|
|
|
Advanced spectroscopic characterization of quantum dot ensembles
Greben, Michael ; Valenta, Jan (advisor) ; Linnros, Jan (referee) ; Vácha, Martin (referee)
Title: Advanced spectroscopic characterization of quantum dot ensembles Author: Michael Greben Department: Department of Chemical Physics and Optics Supervisor of the doctoral thesis: Prof. Jan Valenta, Ph.D. Abstract: Semiconductor quantum dots (QDs) are small crystallites whose sizes (of the order of nm) cause spatial confinement of carriers in all 3 dimensions. As result, QDs often reveal very different physical properties in comparison with their bulk counterparts. From the optical point of view, the broadening of bandgap with QD-size shrinking is particularly interesting. It is a purely quantum mechanical effect that results from quantum confinement (QC), i.e. dimensional limitations of excitons. A strong spatial confinement leads to a relaxation of momentum (Heisenberg uncertainty principle), consequently, larger overlap of the wave-functions of carriers results in significant increase of probability of radiative recombination. Therefore ensembles of QDs are promising candidates for new generations of photonic and photovoltaic devices. This PhD thesis is primary focused on detailed spectroscopic characterization of ensembles of direct (PbS) and indirect (Si) semiconductor QDs in both colloidal (toluene) and matrix-embedded (oxide or oxinitrides multilayers) forms. The oleic- acid capped PbS QDs were...
|
|
| |
|
|
Advanced spectroscopic characterization of quantum dot ensembles
Greben, Michael ; Valenta, Jan (advisor)
Title: Advanced spectroscopic characterization of quantum dot ensembles Author: Michael Greben Department: Department of Chemical Physics and Optics Supervisor of the doctoral thesis: Prof. Jan Valenta, Ph.D. Abstract: Semiconductor quantum dots (QDs) are small crystallites whose sizes (of the order of nm) cause spatial confinement of carriers in all 3 dimensions. As result, QDs often reveal very different physical properties in comparison with their bulk counterparts. From the optical point of view, the broadening of bandgap with QD-size shrinking is particularly interesting. It is a purely quantum mechanical effect that results from quantum confinement (QC), i.e. dimensional limitations of excitons. A strong spatial confinement leads to a relaxation of momentum (Heisenberg uncertainty principle), consequently, larger overlap of the wave-functions of carriers results in significant increase of probability of radiative recombination. Therefore ensembles of QDs are promising candidates for new generations of photonic and photovoltaic devices. This PhD thesis is primary focused on detailed spectroscopic characterization of ensembles of direct (PbS) and indirect (Si) semiconductor QDs in both colloidal (toluene) and matrix-embedded (oxide or oxinitrides multilayers) forms. The oleic- acid capped PbS QDs were...
|
|
|
Study of metallothionein conjugates by electromigration methods
Johaníková, Klára ; Zemánková, Kristýna (referee) ; Vaculovičová, Markéta (advisor)
The bachelor thesis deals with the study of conjugates of metallothionein (MT) with fluorescent nanoparticles using electromigration methods. Particular attention is paid to the preparation and characterization of quantum dots (QDs) and their bioconjugates with the metallothionein protein, whose separation conditions (concentration and suitable electrolyte pH) have been carefully optimized. For the preparation of QDs, the action of UV radiation for one type of quantum dots and the thermal action of high temperature for the second type of species were used. Conjugation of QDs to the protein was performed via a carbodiimide-zero-length crosslinker (EDC / sulfo-NHS), which activates carboxyl groups and allows bioconjugation of the ligand by covalent bonding. The resulting bioconjugates were studied by capillary electrophoresis with absorption detection ( 214).
|
|
|
Luminiscenční nanočástice pro bioanalytické aplikace =: Luminescent nanoparticles for bioanalytical applications /
Vaněčková, Tereza
The dissertation thesis entitled Luminescence nanoparticles for bioanalytical applications deals with the use of optical nanomaterials in life sciences. An overview of the commonly used luminescent nanoprobes is provided together with their advantages over commonly used organic dyes or fluorescence proteins. Next, surface modifications and biofunctionalization of nanoparticles with targeting moieties are discussed. Molecularly imprinted polymers are introduced as an alternative surface modification enabling biorecognition. Finally, theoretical part is concluded with recent examples of the luminescent nanoparticles in bioanalytical and imaging applications. The scientific results of the Ph.D. candidate are presented in the form of 2 review articles and 3 research articles in the peer reviewed journals.
|
|
|
Studium vzniku a vlastností fluorescenčně aktivních nanočástic
Prášková, Markéta
Nanoparticles belong to the future of mankind, both in the scientific world and objects of everyday life. Their potential is apparent not only in the diagnostics and treatment of various diseases, but also in engineering or informatics. Nanoparticles with fluorescent properties can be used for the fluorescent labelling of molecules for the monitoring of various biological processes, tracking of the fate of administered drug in cells or whole organisms, or for the construction of biosensors. Presented work deals with two types of nanoparticles - liposomes and quantum dots (QDs). In this work, the properties of liposomes with encapsulated fluorescent active drug doxorubicin were evaluated. Moreover, QDs of various sizes were characterized, synthesized by microwave synthesis and biosynthesis. The nanoparticles were analysed by their optical properties, zeta-potential and toxicity propertis. Other methods, such as gel electrophoresis, electrochemistry, liquid chromatography, mass spectrometry or Raman spectroscopy were also used in this work.
|
|
|
New technique on a chip for rapid detection of biological materials
Pejović Simeunović, Jelena ; Foret,, František (referee) ; Táborský,, Petr (referee) ; Hubálek, Jaromír (advisor)
Tato práce navrhuje techniku separace a detekce na čipu pro kvantové tečky (QD, „quantum dots“) konjugované s různými proteiny, za účelem sledování vlivu vazebného činidla na potlačení intenzity uorescence QD způsobené konjugací s proteinem a za účelem provedení multianalytické imunoanalýzy k identifikaci malých množství daného proteinu. Za optimálních podmínek byly biokonjugované QD úspěšně odděleny od těch nezkonjugovaných během 10 minut. Částice a cílové roztoky byly smíchány a detekce na čipu byla provedena za pomoci zařízení vyvinutého v naší laboratoři. Byl použit pouze jeden zdroj excitačního světla v kombinaci s několika filtry pro různé emisní vlnové délky. Fluorescence emitovaná dvěma typy konjugovaných QD mohla být poté zaznamenána současně, protože QD emitovaly světlo na různých vlnových délkách, ačkoliv byly excitovány při stejné vlnové délce. Smícháním dvou typů QD biokonjugovaných se dvěma druhy proteinů a protilátek jsme dokázali detekovat imunokomplexní píky s různými plochami. Plocha pod píkem závisela na koncentraci QD a antigenů, na postupu reakcí protilátka–antigen a ukázalo se, že je lineárně korelována s koncentrací antigenu. Ukázali jsme, že kapilární elektroforéza QD na čipu může být použita jako citlivá technika pro detekci biologických molekul. Hlavními výhodami této metody jsou jednoduchost, malé požadavky na objem vzorku i činidla a také vysoká účinnost separace.
|
|
|
New pathways to plasmonic nanoparticle assembling into 2D and 3D hybrid active systems for SERS of graphene and SERS, SERRS and GERS + SERS of aromatic molecules
Gajdošová, Veronika ; Vlčková, Blanka (advisor) ; Němec, Ivan (referee) ; Michl, Martin (referee)
In the first part of the Thesis, a new type of active system for SERS and SERRS of hydrophobic molecules, namely a 3-dimensional (3D) nanosponge aggregate with incorporated hydrophobic molecules has been developed, and tested by fullerene C60 and hydrophobic free- base tetraphenylporfine (H2TPP). The SERS and SERRS (surface enhanced /resonance/ Raman scattering) limits of detection (LODs) of C60 at four excitation wavelengths spanning the visible spectral region were found to be by one order of magnitude lower than in the reference system, which mimics the previously reported ways of utilization of Ag nanosponges as substrates for SERS and SERRS. The superiority of the newly developed sample is attributed to the efficient localization of the hydrophobic molecules into hot spots in 2D fractal aggregates of Ag nanoparticles (NPs). Diprotonation of H2TPP during the procedure using HCl as the preaggregation agent has been eliminated by employment of NaCl. On the other hand, investigation of the mechanism of H2TPP protonation during the former preparation procedure opened a possibility to employ Ag nanosponge aggregate as nanoreactor. In the second part of the Thesis, 2D assemblies of AgNPs were found to be better substrates for SERS of single layer graphene (SLG) than the 3D ones. In particular, the 2D...
|
|
|
Analysis of biotic matrices by Laser-Induced Breakdown Spectroscopy
Střítežská, Sára ; Karel,, Novotný (referee) ; Modlitbová, Pavlína (advisor)
This bachelor thesis discusses the possibility of using Laser-induced breakdown spectroscopy (LIBS) for biological application. The LIBS method was used to determine spatial distribution of a chosen element, cadmium, in selected model plant, white mustard (Sinapis Alba L.) The exposure took place in hydroponic conditions and lasted for 72 hours, therefore it was an acute toxicity test. Three different sources of cadmium were tested, mercaptopropionic acid stabilized quantum dots (CdTe QDs), silica coated quantum dots (CdTe/SiO2 QDs), and cadmium chloride, which was used as a positive control. After the exposure and before determination of the spatial distribution of cadmium in plants, a significant portion of the thesis was devoted to optimization of selected parameters of LIBS measurement (laser energy and gate delay). Subsequently, the cadmium distribution was determined and the obtained results were discussed with regards to the tested substance, its stability and its tendency to aggregate/agglomerate in water medium in contact with testing organism during exposure. In the final part of the work, the suitability of the LIBS method for the determination of spatial distribution of chosen contaminants was evaluated.
|
guest ::
