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Golden nanosystems for molecular detection of biological samples using Surface-Enhanced Raman Spectroscopy
Kebabová, Mona ; Bernátová,, Silva (referee) ; Samek, Ota (advisor)
Citlivá a přesná detekce analytů nebo bakterií je klíčovým krokem při identifikaci nebezpečných sloučenin či patogenů, ať už v oblasti medicíny nebo bezpečnosti potravin. Využití pokročilých metod, jako je povrchově zesílená Ramanova spektroskopie (SERS), by pomohlo identifikovat bakterie nebo analyty s vysokou citlivostí a přesností. Tato diplomová práce zaměřuje na syntézu a charakterizaci kovových nanočástic (NČ) pro citlivou detekci bakterií pomocí SERS, přičemž NČ zvyšují Ramanův signál lokalizovanou plazmonovou rezonancí. Nejdříve jsou diskutovány nejnovější přístupy k syntéze plazmonických nanostruktur. Hlavní část experimentální práce tvoří syntéza zlatých (Au) a stříbrných (Ag) NČ, jejich charakterizace a využití pro analýzu chemických a biologických vzorků pomocí SERS. Nanočástice byly použity pro detekci molekul barviv (rhodamin 6G a methylenová modř), a patogenních bakterií Staphylococcus aureus a Escherichia coli, adsorbovaných na povrch syntetizovaných NČ. Dále byl replikován vysoce specifický sendvičový imunotest detekovatelný pomocí SERS. Tato metoda umožňuje rychlou a přesnou detekci patogenů. Na povrch nanočástic se pomocí interakce avidin-biotin vážou protilátky, které slouží k zachycení bakterií (E. coli a S. aureus). Ačkoli dílčí kroky protokolu byly úspešné, ověření funkce sendvičového imunotestu se doposud nezdařilo, pravděpodobně kvůli změnám v použitých chemikáliích a protilátkách. Kroky nezbytné k replikaci protokolu budou provedeny v budoucnu.
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Gold nano systems for the detection of molecules using surface-enhanced Raman scattering (SERS)
Benešová, Markéta ; Bernatová,, Silvie (referee) ; Skoumalová, Petra (advisor)
This diploma thesis was focused on the production and use of gold nanosystems to enhance Raman scattering. Metal nanoparticles, when interacting with electromagnetic radiation, form so-called localized plasmons, which can enhance Raman scattering. SERS (surface-enhanced Raman scattering) is a non-destructive analytical technique used in this work to measure the concentration and chemical changes in the rhodamine B molecule. Rhodamine B was subjected to photocatalytic degradation using two types of photocatalysts before measurement: TiO2-(H) and TiO2-(H)-Ag. The quantitative dependence of rhodamine B photodegradation on the presence of photocatalysts in a given period time was sought using photodegradation processes. The results of measurements using the SERS method were compared with the results obtained by UV-VIS spectroscopy. From the measured data, it was found that the photocatalysts significantly accelerate the photodegradation processes, because the Raman signal of rhodamine B decreased, while the signal decrease was most pronounced for the catalyst with added TiO2-(H)-Ag, less prominent but still statistically significant signal decrease was observed for the TiO2-(H) catalyst. In the control sample without the addition of photocatalyst, no decrease in signal was observed. In the next phase of the thesis, a sandwich immunoassay was designed that uses SERS to detect E. coli bacteria or other specific microorganisms in the sample. The first component of the sandwich immunoassay is gold nanoparticles, which carry a so-called Raman reporter, which has a clear Raman response in the spectrum, and gold nanoparticles amplify this signal, and antibodies, thanks to which the particles specifically bind to the microorganism. Another component is either gold layered slides or magnetic nanoparticles, which are modified with antibodies and serve to immobilize microorganisms. This system can be a fast and very accurate way to identify a given microorganism in a sample.
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Gold nano systems for the detection of molecules using surface-enhanced Raman scattering (SERS)
Benešová, Markéta ; Bernatová,, Silvie (referee) ; Skoumalová, Petra (advisor)
This diploma thesis was focused on the production and use of gold nanosystems to enhance Raman scattering. Metal nanoparticles, when interacting with electromagnetic radiation, form so-called localized plasmons, which can enhance Raman scattering. SERS (surface-enhanced Raman scattering) is a non-destructive analytical technique used in this work to measure the concentration and chemical changes in the rhodamine B molecule. Rhodamine B was subjected to photocatalytic degradation using two types of photocatalysts before measurement: TiO2-(H) and TiO2-(H)-Ag. The quantitative dependence of rhodamine B photodegradation on the presence of photocatalysts in a given period time was sought using photodegradation processes. The results of measurements using the SERS method were compared with the results obtained by UV-VIS spectroscopy. From the measured data, it was found that the photocatalysts significantly accelerate the photodegradation processes, because the Raman signal of rhodamine B decreased, while the signal decrease was most pronounced for the catalyst with added TiO2-(H)-Ag, less prominent but still statistically significant signal decrease was observed for the TiO2-(H) catalyst. In the control sample without the addition of photocatalyst, no decrease in signal was observed. In the next phase of the thesis, a sandwich immunoassay was designed that uses SERS to detect E. coli bacteria or other specific microorganisms in the sample. The first component of the sandwich immunoassay is gold nanoparticles, which carry a so-called Raman reporter, which has a clear Raman response in the spectrum, and gold nanoparticles amplify this signal, and antibodies, thanks to which the particles specifically bind to the microorganism. Another component is either gold layered slides or magnetic nanoparticles, which are modified with antibodies and serve to immobilize microorganisms. This system can be a fast and very accurate way to identify a given microorganism in a sample.
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