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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.
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Plasmon enhanced photoluminescence
Édes, Zoltán ; Kůsová,, Kateřina (referee) ; Šikola, Tomáš (advisor)
Diplomová práce se zabývá fotoluminiscencí polovodičových materiálů zesílené plazmonovými polaritony. Je popsána základní teorie interakce mezi lokalizovanými povrchovými plazmonovými polaritony a fotoluminiscenčními látkami. Dva mechanismy, které mohou vést k fotoluminiscenci zesílené plazmonovými polaritony jsou diskutovány. Následně je popsán návrh aparatury pro měření fotoluminiscence a způsob její realizace. Funkčnost aparatury je ověřena měřením fotoluminiscenčních spekter objemového GaN, nanokrystalického Si a CdTe kvantových teček. Nakonec je zkoumána metoda přípravy vzorků sestávajících z kovových nanokuliček a fotoluminiscenčně aktívních CdTe kvantových teček.
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Strong coupling in plasmonic antennas
Beneš, Adam ; Křápek, Vlastimil (referee) ; Kejík, Lukáš (advisor)
Localized surface plasmons generated by ilumination of metallic nanostructures focuses light into nanoscale regions where it can interacts with other nanoobjects nearby. This bachelor's thesis focuses on fabrication of plasmonic antennas suitable for strong coupling with non-conductive or semiconductor nanostructures such as dye molecules or quantum dots in visible range. At first we put forward some theoretical background and summary of strong coupling systems. In the main section we describe process of fabrication and characterization of plasmonics antennas as well as deposition methods for emitors. Lastly, we study interactions in our systems using spectroscopic measurements and strong coupling is realized using FDTD simulations.
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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.
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Biosensors based on modified nanoparticles
Mihajlović, Ana ; Majzlíková, Petra (referee) ; Hubálek, Jaromír (advisor)
This thesis deals with biosensors based on modified semiconductor quantum dots(AIIBIII). This work is divided into three main parts. The first deals with the theory necessary for the use of QDs in bioapplications, there are described methods of synthesis, application and bioconjugation of QDs . In the practical part, GSH-CdTe, CdTe-MPA and TGA-CdTe quantum dots were prepared by direct colloidal synthesis. These quantum dots were then modified by EDC in order to increase affinity to BSA (bovine serumalbumin). For the analysis of samples were used Infinite M200,Tecan and HORIBA Jobin Yvon FluoroLog detector Quanta . The third part is the summary of results and work achievements.
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Synthesis of core/shell quantum dots for diagnostics
Mihajlović, Ana ; Vaculovičová, Markéta (referee) ; Pekárková, Jana (advisor)
This thesis deals with biosensors based on modified semiconductor core/shell quantum dots (QDs) for diagnosis. The work is divided into four main parts. The first one discusses the theory required for the use of QDs in bioaplications, there are described methods of synthesis, modification, application and bioconjugation of QDs. In the experimental part, CdTe/ZnS QDs with core/shell structure were prepared, in which the core was modified by MPA, GSH and TGA. In the next step, these QDs were further modified using CDI, EDC and NHS as mediators in order to increase affinity to BSA (bovine serum albumine) and IgG (imunoglobuline G). Prepared conjugates were characterized by fluorescence spectroscopy (Infinite M200Pro, Tecan) and capillary electrophoresis (Agilent 7100).
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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.
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Nanostructures and Nanosensors
Křečková, Jitka ; Haupt, Jaromír (referee) ; Ellederová, Eva (advisor)
Cílem této bakalářské práce je představit využití nanotechnologie ve snímacích zařízeních, poskytnout základní informace o nanotechnologii, včetně popisu a klasifikace nanostruktur, a zdůraznit možnost vylepšení výkonu senzoru prostřednictvím nanotechnologie. Tato práce se také zabývá využitím nanodrátu jako součástí senzoru s konfigurací unipolárního tranzistoru. Kvantové tečky a jejich využití v optické detekci jsou také zahrnuty v této práci. Výhody i současná omezení nanosenzorů jsou shrnuty v závěru práce.
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Biomolecules influence on optical properties of deposited quantum dots
Roček, Vojtěch ; Mravec, Filip (referee) ; Drbohlavová, Jana (advisor)
This thesis is focused on creation of quantum dots of titanium dioxide with help of nanoporous alumina mask and on funcionalization of their surface in purpose to increase the quantum yield of its. This thesis is devided into 6 main chapters, where the glossary topic of quantum dots creation process and ways of practical usage of these quantum dots are made. The theoretical knowledge is used for creation of titanium dioxide quantum dots, which was deposited by gold and funcionalized with glutation. The results are shown and discussed in the end of this work.
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