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Dissertations / Theses on the topic 'Biosenzors'
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1
Pavlásková, Lucie. "Biosenzory na bázi funkcionalizovaného grafenu." Master's thesis, Vysoké učení technické v Brně. Fakulta strojního inženýrství, 2021. http://www.nusl.cz/ntk/nusl-443227.
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V této práci byl demonstrován grafenový polem řízený transistor (GFET) jako platforma pro detekci glukózy. Sukcinimidyl ester pyrenbutanové kyseliny (PSE) sloužící jako nosič a enzym glukóza oxidáza (GOx) byly úspěšně použity k funkcionalizaci grafenového kanálu ve FE transistoru. Enzym GOx byl imobilizován na kanálu pro glukózovou detekci, jelikož indukuje selektivní katalytickou reakci glukózy. Proces funkcionalizace byl charakterizován pomocí Ramanovy spektroskopie a Atomární silové mikroskopie (AFM). Vyrobený biosenzor na bázi grafenu umožnil elektrickou detekci glukózy ve dvou různých uspořádáních. V uspořádní FET prostřednictvím posunu Diracova bodu ve voltampérové charakteristice, jakož i v nastavení pro kotinuální monitorování v reálném čase prostřednictvím změny odporu grafenového kanálu. Tato studie naznačuje, že grafen je slibným materiálem pro vývoj nanoelektronických biosenzorů včetně aplikací pro monitorování hladiny glukózy.
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Přítulová, Marie. "Funkční nanočástice pro plasmonické biosenzory." Master's thesis, Vysoké učení technické v Brně. Fakulta chemická, 2016. http://www.nusl.cz/ntk/nusl-240594.
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This thesis aims to prepare functional gold nanoparticles (AuNPs) and use them in conjunction with a surface plasmon resonance (SPR) biosensor for highly sensitive detection of carcinoembryonic antigen (CEA). In this work, preparation of colloidal AuNPs was investigated and a three-step synthesis was optimized to yield spherical nanoparticles with a diameter of about 100 nm and smooth surface. The synthesized AuNPs were functionalized by a self-assembled monolayer of carboxy-PEG alkanethiols and streptavidin and characterized by UV/VIS spectroscopy and -potential method. Finally, the functionalized AuNPs were employed in sandwich assay for the sensitive detection of CEA and it was demonstrated that they can enhance sensor response to CEA by a factor of 100 compared to the direct detection of CEA.
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Matula, Tomáš. "Elektrochemický biosenzor pro detekci glukózy." Master's thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2016. http://www.nusl.cz/ntk/nusl-241997.
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This thesis pays attention to electrochemical sensors for the detection of glucose. The theoretical part deals with the technology of thick film, their application and firing. It is further described electrochemistry, electrodes used in electrochemistry and analytical methods based on electrochemical reaction. End of theoretical part deals with electrochemical biosensors for detection glucose using special electrodes based on enzyme glucose oxidase. In the experimental part is processed amperometric measure for detection glucose using screenprinted sensors S10 and S8 with immobilized enzyme glucose oxidase. Next was studied effect of plasma, dissolved potassium chloride, storage, ascorbic acid and the reproducibility of the sensors.
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Kynclová, Hana. "Charakterizace nanostrukturovaných elektrod pro elektrochemické biosenzory." Master's thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2012. http://www.nusl.cz/ntk/nusl-219511.
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Nowadays it is attached to a major effort to study applications of nanoparticles in biosensors technology. We studied the effect of gold nanoparticles on the surface of the electrodes by Electrochemical Impedance Spectroscopy method and Cyclic Voltammetry. For impedance measurements was proposed substitute electrical model and cyclic voltammetry method was used to determine the electroactive surface of electrodes.
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Tripský, Andrej. "Návrh, výroba a testování grafenových biosenzorů." Master's thesis, Vysoké učení technické v Brně. Fakulta strojního inženýrství, 2020. http://www.nusl.cz/ntk/nusl-417170.
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Pokrok ve vývoji nanotechnologií nám poskytuje dobrou příležitost k vývoji nových špičkových zařízení. Tato práce si klade za cíl vyrobit, popsat a změřit grafenové pH senzory na dvou různých substrátech - polymeru parylenu C a SiO2. Tento pH senzor je prvním krokem ve vývoji nositelné náplasti monitorující stav kůže a možné infekce. Grafen je 2D materiál na bázi uhlíku se zajímavými vlastnosti a nadějnými aplikacemi. Úspěšně jsme provedli dva různé experimenty sloužící k charakterizaci grafenových senzorů a jejich odezvu na různé hodnoty pH. V prvním experimentu jsme použili horní elektrolytické hradlo k určení bodu neutrality (Diracův bod). Druhý experiment popsal změnu rezistence grafenu jako funkce pH. Dále jsme také funkcionalizovali grafen polyanilinem, abychom zlepšili jeho vlastnosti. Prokázali jsme citlivost grafenových senzorů na pH pro oba substráty a objevili jsme několik výzev jako potřebu kontroly iontové síly, experimentů samotných a destrukce grafenu.
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Petrula, Jakub. "Elektrochemický biosenzor pro studium metylace DNA." Master's thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2017. http://www.nusl.cz/ntk/nusl-316815.
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This bachelor’s thesis deals with design and optimalisation of custom biosensor for detection of methylated DNA. Teoretical part explains the mechanism and importance of DNA methylation. Next section describes analytical methods used in connection with DNA methylation and some basic direct and indirect methods of detection. Final part is dedicated to experiment itself, which is divided into several sections. Section one deals witch modification of working electrode and optimalisation of detection method. Second section introduces two different ways of DNA methylation detection. First is based on direct detection and second one on detection through the biosensor. Final part shows determination of methylcytosine from sample based on analysing characteristic attributes of signal and numeric algorithm based on curve fitting.
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Dršata, Martin. "Plazmonické biosenzory založené na zvýšené optické transmisi." Master's thesis, Vysoké učení technické v Brně. Fakulta strojního inženýrství, 2017. http://www.nusl.cz/ntk/nusl-318371.
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Tato diplomová práce se zabývá rigorózními simulacemi plazmonických biosenzorů založených na jevu zvýšené optické transmise. První část je věnována popisu fyzikálních jevů a poznatků, které tvoří základ pro studium vlastností plazmonických senzorů, a popisu výpočetní metody konečných prvků v časové oblasti, která je využita v této práci. Vlastní výsledky jsou uvedeny v další části, která se zabývá výzkumem citlivosti, rozlišení a dalších charakteristik zvoleného typu plazmonického sensoru, tvořeného sítí kruhových nanoděr v tenké zlaté vrstvě na substrátě nitridu křemíku, v závislosti na řadě jeho geometrických parametrů. Tyto závislosti jsou sledovány ve třech různých případech, a to senzoru umístěného ve vakuu, ponořeného ve vodě a v případě kdy je na zlatém povrchu umístěna tenká dielektrická vrstva, která reprezentuje přítomnost biomolekul uchycených na povrchu senzoru.
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Sedláček, Jiří. "Biosenzor halogenovaných látek jako přístroj využívající fluorescenční metodu." Master's thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2010. http://www.nusl.cz/ntk/nusl-218678.
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This master thesis deals with design and construction of device for optic biosensor. The device are using for detected halogen compound in the water. For excitation testing sample is used LED diode about wave length 590 nm. Photocounter is used for detection of the emitted light after fluorescence Such as fluorescent marker are 5(6)-carboxynaphthofluorescein. The thesis is divided into three head parts. The first part includes the theory needed for realization; the second part takes in detail description of the construction and function of the device and the last part describes measurements with the device.
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Křepelka, Pavel. "NÁVRH OPTOVLÁKNOVÉHO BIOSENZORU SE SPEKTRÁLNÍ ANALÝZOU V BLÍZKÉ INFRAČERVENÉ OBLASTI." Doctoral thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2019. http://www.nusl.cz/ntk/nusl-409090.
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This thesis deals with a measurement and interpretation of NIR spectra of bacterial cells and design of biosensor using this analytical technique. In the first chapter, there is introduction of current state of knowledge in the field of NIR spectroscopy in microbiology and technology of fiber optic biosensors. The summary of this chapter shows that NIR is a suitable technique for direct molecular analysis of bacteria, but it suffers from low sensitivity and insufficient interpretation of bacterial spectra. In the next part of the thesis, there is a theoretical background of spectral analysis techniques and technology of fiber optic sensors. In the practical part of this work, there is suggested the elimination of disadvantages of NIR spectroscopy in microbiology by a series of experiments used for interpretation of NIR spectra of bacteria and design of fiber optic sensor to increase sensitivity of this technique. In this work, spectral regions important for the identification of bacterial strains were determined and partially interpreted and the sensor for bacterial analysis capable of classifying strains based on 105 captured cells was designed. Therefore, the objectives of this work were fulfilled.
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Láznička, Jan. "Analýza spolehlivosti měření látek pomocí biosenzorů - návrh robustní metody stanovení limitu detekce." Master's thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2008. http://www.nusl.cz/ntk/nusl-217622.
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Detection limit of any method is affected by device (electronics, noise), evaluation, software (timing inaccuracy, rounding mistakes, errors determination of parameters using measured values by numerical methods) and by a chemical reaction (diluting errors, the reaction mixture design, chemical interferents, temperature, the accuracy of pipetting). In this work was proposed a metod for determination of detection limit for measuring of inhibitors of acetylcholine esterase by biosensor analysator of toxicity (BTA). The analysator was developed by BVT Technologies a.s. in 2004. To determine the exact detection limit a calibration of output signal was done. Presently the analysator is able to detect only presence of toxins, not their concentration. Not least are mentioned all founded errors of software, which are continuously eliminated.
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Šálek, Petr. "Příprava a charakterizace magnetických nosičů z hypersíťovaných polystyrenových mikročástic a jejich použití v biosenzoru." Doctoral thesis, Vysoké učení technické v Brně. Fakulta chemická, 2012. http://www.nusl.cz/ntk/nusl-233348.
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With the aim to develop and characterize a functionalized highly magnetic polymer carrier of micrometer size and of a narrow particle size distribution that will be suitable for biological application, hypercrosslinked microspheres were prepared. Simultaneously, the relation between structure and properties of product was observed. Condition of dispersion polymerization were optimized to obtain starting monodisperse poly(styrene-co-divinylbenzene) [P(St-DVB)] microspheres. The P(St-DVB) microspheres of different degree of crosslinking were prepared and effect of some polymerization parameters such as type of solvent, initiator, concentration and mode of DVB addition on morphology, size and particle size distribution were investigated. The starting microspheres were hypercrosslinked to obtain microporous inner structure. Hyperosslinked particles had very large specific surface area (> 1000 m2/g) and a high content of micropores (ca. 0.6 ml/g). First, P(St-DVB) microspheres were chloromethylated using three different chloromethylation agents to regulate their porous properties. Hypercrosslinking was achieved by the addition of stannic chloride as a catalyst and by increasing a temperature. The hypercrosslinked microspheres were then functionalized with sulfo- or aminogroups. The functional groups captured precipitated iron oxide inside the porous structure of the microspheres and also served as a reactive site for intended immobilization of the protein. A solution of ferrous and ferric chloride was imbibed under vacuum into the porous structure and the iron oxide was precipitated by an aqueous ammonia solution. Finally, the magnetic functionalized hypercrosslinked micropsheres were integrated into a biosensor for qualitative detection of ovalbumin.
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Janíček, Zdeněk. "Magnetické částice jako reverzibilní nosič pro enzymové elektrody." Master's thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2012. http://www.nusl.cz/ntk/nusl-219744.
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Master’s thesis contains information about the enzymes, biosensors, enzyme biosensors and magnetic particles. Cholinesterasa (ChE) is the generally name for the two related enzymes, mutually differing appearance and biological functions. Acetylcholinesterasa (AChE) is necessary for the transmission of nervous excitement. AChE is located at cholinergic synapses, where is the hydrolysis of the neurotransmitter acetylcholine and this termination by the nervous excitement. Butyrylcholinesterasa (BChE) is related to AChE and it is present in serum. Experimental part was focused on electrochemical biosensors with cholinesterase enzyme using magnetic particles for reversible reconstruction layer. Cholinesterase immobilization was carried out on the magnetic particles by covalent binding after glutaraldehyde activated. The measurement of activity take place with acetylthiocholin as a substrate in a flow system, magnetic particles were captured before the platinum electrode and produced by thiocholin is detected amperometric. The aim was to measure inhibition of AChE. Significant inhibition ChE is caused by certain drugs and pesticides based on organophosphates. There was tested by inhibition of AChE pesticide dichlorvos. Emphasis was on finding possible experimental conditions for the creation of the automated procedure to measure the AChE activity, which was based on auto-injector and pumps of Gilson, together with a 735 programme for the definition of the working configuration, preparation of measurement scripts and implementation of application methods.
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Ehlich, Jiří. "Optimalizace tiskových metod přípravy organických polovodivých vrstev." Master's thesis, Vysoké učení technické v Brně. Fakulta chemická, 2017. http://www.nusl.cz/ntk/nusl-316199.
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Electrophysiological biosensors enables a novel way to measure electrical activity of biological structures both in-vitro and in-vivo and represents valuable alternative to current cellular activity measuring methods. Within this work we will be focusing on development of organic semiconductor (PEDOT:PSS) based Organic Electrochemical Transistors (OECTs) and optimization of material printing methods used in their development. These transistors are meant to be able to transfer electrochemical signals within the cell membrane to electrical signal. Such sensors should be used for cytotoxicity testing of chemicals and potential drugs on cardiomyocytes. Main benefits of OECTs are in their higher sensitivity thanks to their ability to locally amplify electric signals, better noise-signal ratio and outstanding biocompatibility. Their development is undemanding and inexpensive due material printing methods and materials processable at room temperatures.
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Kejík, Lukáš. "Použití kovových a polovodičových nanostruktur pro biodetekci." Master's thesis, Vysoké učení technické v Brně. Fakulta strojního inženýrství, 2015. http://www.nusl.cz/ntk/nusl-232130.
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The master’s thesis deals with two applications of gold discoidal nanostructures exhibiting plasmon resonance for biodetection. The first approach considers the detection of changes in the phase on plasmonic antennas using coherence-controlled holography microscope. It was found that the steepness of the phase is increasing with the illumination wavelength when plasmon resonance is excited in larger antennas. The sensitivity of the phase to refractive-index changes of the surrounding media was observed when the largest response was given by antennas in resonance with wavelength of illumination. Next part deals with plasmon resonance detection by means of optical spectroscopy combined with voltametry which characterizes the electrochemical activity. Changes in resonance wavelength induced by the presence of SSC buffer were observed, although this influence seems to diminish in time. Conducted experiments have also shown that oxygen-plasma cleaning is not suitable for sample surface cleaning because of oxidation of metals including gold as well.
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Ondruch, Vít. "Analýza signalů tlustovrstvých ampérometrických senzorů a jejich použití pro měření a charakterizaci enzymů." Doctoral thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2009. http://www.nusl.cz/ntk/nusl-233494.
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V práci je popsán princip synchronní detekce (SD), který byl uplatněn při měření s biosenzory. Metoda SD umožňuje dosažení výrazně lepšího poměru signálu k šumu, vyššího limitu detekce a celkové zlepšení robustnosti měření. Uplatnění SD při měření s biosenzory umožní zlepšit analýzu jeho odezvy a umožní odstranění nežádoucích interferencí nebo šumů, které mohou být způsobeny například mícháním roztoku, elektromagnetickými vlivy nebo parazitními proudy. SD také umožňuje rozložit získaný signál na odezvu stimulace a na dlouhodobý signál jiného procesu, a dále také identifikovat jevy druhého řádu. Pro identifikaci stimulačního signálu ve výstupním signálu měření byl na základě lineárního statistického modelu vyvinut specializovaný software. SD byla ověřena na modelovém případu výstupního signálu biosenzoru s aplikovaným komplexem fotosystému II (PSII) a jeho odezvě na stimulaci světlem. Odezva PSII se řídí kinetikou prvního řádu a může být také ovlivněna inhibitory. Kinetické konstanty vazby herbicidu na PSII závisí lineárně na koncentraci herbicidu. To umožňuje jejich měření také při nízkých koncentracích herbicidu.
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Bráblíková, Aneta. "Mikroelektrodová pole pro bioelektroniku." Master's thesis, Vysoké učení technické v Brně. Fakulta chemická, 2019. http://www.nusl.cz/ntk/nusl-401911.
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Organic electronic biosensors are developed as suitable devices that can transform electrochemical processes within the cell membrane into an electronic signal and enable to measure electrical activity of excitable cells and tissues both in vitro and in vivo and thus represent valuable alternative to current cell monitoring methods. In this work we focus on the fabrication of electrophysiological sensors based on organic semiconductors printed by the material printing method. Microelectrode arrays (MEAs) are active components of the device, which can monitore cellular activity and above that stimulating cells with electrical pulses. The proposed platform should be used for cytotoxicity of potential drugs especially on cardiac cells (cardiomyocytes). The experimental part focus on specific production processes of platforms, which were prepared in the laboraty with emphasis on biocompatibility and conductivity of device.
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Hrdý, Radim. "Techniky přípravy elektrod s nanostrukturovaným povrchem a jejich charakterizace." Doctoral thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2013. http://www.nusl.cz/ntk/nusl-233608.
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Nowadays, nanostructures fixed on solid substrates and colloidal nanoparticles permeate through all areas of human life, in area of sensors and detection as well. This dissertation thesis deals with the fabrication of nanostructures on the surface of planar electrodes via self-ordered nanoporous template of aluminum trioxide. The nanofabrication, as one of many possible techniques, is used to increase the active surface area of electrodes by creating unique surface types with specific properties. These electrodes are very perspective in the applications, such as biomolecules electrochemical detection and measurement. The transformation of aluminum layer into non-conductive nanoporous template in the process of anodic oxidation is a fundamental technique employed to obtain the array of nanostructures in this thesis. The fabrication of high quality nanoporous membranes with narrow pore size distribution on various types of metallic multilayers is one of the key experimental parts in this work. Several problems associated with the production of the thin-film systems, including the dissolving the barrier oxide layer, are discussed and solved. Another part of this work deals with the use of nanoporous membrane as a template for the production of metallic nanostructures via electrochemical metal ions deposition directly into the pores. The obtained nanostructures as nanowires, nanorods or nanodots are characterized by the scanning electron microscopy and energy-dispersive or wavelength X-ray spectroscopy. The electrode surface, modified by gold nanostructures suitable for the detection of biomolecules, has been chosen for the electrochemical measurements, due to the gold biocompatibility. The nanostructured electrodes were characterized by electrochemical impedance spectroscopy and cyclic voltammetry. The effect of nanostructured surface geometrical parameters, including the size of the electrochemically active area, on the results of electrochemical measurements has been observed and compared to flat gold electrodes. Two model biomolecules, namely guanine and glutathione, have been chosen for the study of potential application of these nanostructures in biosensors.
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Omasta, Lukáš. "Printed Biosensor Based on Organic Electrochemical Transistor." Doctoral thesis, Vysoké učení technické v Brně. Fakulta chemická, 2019. http://www.nusl.cz/ntk/nusl-402112.
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Organické elektronické zariadenia sú vyvíjané ako vhodné riešenia senzorov pre bioelektroniku, a to najmä kvôli dobrej biokompatibilite organických polovodičov v nich použitých. Takzvané biosenzory dokážu premeniť elektrochemické procesy na elektronický signál. Matrica takýchto biosenzorov môže simultánne skenovať množstvo biologických vzoriek, alebo rôznych tkanív v živých systémoch. Aktívnou súčasťou zariadenia je organický elektrochemický tranzistor (OECT). V tejto práci je diskutovaný teoretický rámec fungovania takéhoto zariadenia, jeho elektrická charakterizácia, aplikácia v biosenzoroch na báze buniek, spôsoby výroby a aktuálnym stavom techniky v oblasti organickej elektroniky. Experimentálna časť obsahuje konkrétne výrobné postupy vývoja OECT zariadení, ktoré boli použité v našom laboratóriu. Hlavný dôraz sa kladie na schopnosť vyrobených zariadení detekovať reakciu a monitorovať stimuláciu elektrogenných buniek. Za týmto účelom boli vyvinuté matice mikroelektródových OECT zariadení založených na polovodivom polyméri PEDOT:PSS. Tieto boli vyrobené s využitím bežnými tlačiarenských techník (atramentová tlač a sieťotlač) spolu so štandardnými litografickými postupmi. Najnovšie nami vyvinuté zariadenia dosahujú najväčšieho zosílením signálu, g = 2,5 mS a časovú konštantu t = 0,15 s. Tieto zariadenia sú porovnateľné, často dokonca lepšie ako niektoré iné najmodernejšie a plne litograficky pripravené senzory.
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Marková, Aneta. "Vývoj senzorické platformy pro studium fyziologických funkcí buněk." Master's thesis, Vysoké učení technické v Brně. Fakulta chemická, 2018. http://www.nusl.cz/ntk/nusl-377318.
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The aim was to develop a sensing platform on the base of organic electrochemical transistor (OECT). The focus was on the preparation of proper electrode system and on optimalization of properties of thin layer of organic semiconductor. As a base, commercial glass substrates with integrated indium-tin oxide electrodes were chosen. Thin layers were prepared from organic semiconductor poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) by spin-coating. Four formulations of material were studied. Layers with different thickness were prepared and the dependence of transconductance on the thickness of the layer and ratio of width and length was observed. The degradation of electrode system was solved by galvanic plating with gold. Attention was also paid to modifications to PEDOT: PSS. It has been found that the optimal layer thickness for use in sensors is approximately 150 nm. By reducing the series resistance by using a silver paste, the transconductance of 23 mS was obtained for the Ink 2, for the Ink 3 the transconductance was 44 mS. Sensoric platforms with these transconductances can be used for detection of physiological functions of electrogenic cells, e.g. cardiomyocytes.
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Peksa, Vlastimil. "Designing and testing of new metal nanosubstrates for biomolecular sensors based on surface-enhanced Raman scattering (SERS) spectroscopy." Doctoral thesis, 2017. http://www.nusl.cz/ntk/nusl-367962.
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Title: Designing and testing of new metal nanosubstrates for biomolecular sensors based on surface-enhanced Raman scattering (SERS) spectroscopy Author: Vlastimil Peksa Department: Institute of Physics of Charles University Supervisor: doc. RNDr. Marek Procházka, Ph.D., Institute of Physics of Charles University Abstract: This experimental methodical work was aimed at the optimization of selected gold and silver substrates and their use in construction of SERS-based biosensors, including following practical application. Several types of substrates, fabricated via a combination of bottom-up techniques on solid surfaces, were tested. The properties of these substrates were examined with probe molecules, namely methylene blue, porphyrins and tryptophan, on a confocal Raman microspectrometer. Obtained findings about the influence of analyte application, objective focusing and internal intensity standard were exploited for optimization of measurement procedures with regard to sensitivity, accuracy and reproducibility. A method for quantitative detection of food dye azorubine (E 122) in commercially available drinks was developed, based on these findings. Its results have shown its potential as a pre-scan method for field application and preliminary testing. Keywords: Metal nanosubstrates, biomolecules,...
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Doležalová, Julie. "Vývoj elektrochemického glutamátového biosenzoru." Master's thesis, 2007. http://www.nusl.cz/ntk/nusl-273427.
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DEVELOPMENT OF A GLUTAMATE ELECTROCHEMICAL BIOSENSOR Julie Doležalová (Department of Analytical Chemistry, Faculty of Pharmacy, Charles University in Prague) In cooperation with: Sofia A.L.P. Gomes, Alberto N. Araújo (Department of Physical Chemistry, Faculty of Pharmacy, University of Porto) L-Glutamic acid and monosodium glutamate are used to enhance flavour in food prepared at home, restaurants, and by the food processing industry [1]. Glutamate also plays an important role in many metabolic processes. Nevertheless its use is being kept under certain limits as some people have experienced adverse effects, such as stiffness and tension, after its ingestion. Moreover, some studies demonstrate its connection with the appearance of neurological manifestations, such as Parkinson's and Alzheimer's disease. The determination of L-glutamic acid is therefore of major importance. In this work a novel L-glutamate biosensor was developed using a dehydrogenase enzyme (L-glutamate dehydrogenase). These redox enzyme requires for its activity nicotinamide (NAD(P)+ /NAD(P)H) coenzyme, which reduced form was electrochemically regenerated by the water-soluble dye, methylene green. Graphite electrodes were modified with a poly(methylene green) thin film by means of methylene green electrochemical polymerization...
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Eisenreichová, Andrea. "Vývoj biosenzoru pro fosfatidylinositol." Master's thesis, 2017. http://www.nusl.cz/ntk/nusl-365118.
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Phosphatidylinositol is a a minor membrane component of eukaryotic cells, however, it plays a crucial role in cell signaling pathways as a precursor for a number of signaling molecules and second messengers. Among the most significant ones are phosphoinositides created by phosphorylation of the hydroxyl groups of phosphatidylinositol at positions 3,4, and 5 of the inositol ring. Despite its significance, the spatial and temporal distribution and dynamics of phosphatidylinositol remains unclear owing mainly to the lack of a specific optical probe (biosensor) to visualize phosphatidylinositol in living cells. Biosensor for inositol phospholipids are based on lipid-binding domains of their effector proteins with high enough affinity and specificity for a given phosphoinositide - but nosuch domain is known for PI. However, an enzyme - phosphatidylinositol-dependent phospholipase C - that specifically recognizes phosphatidylinositol is known. This enzyme catalyzes the hydrolysis of phosphatidylinositol into diacylglycerol and inositol 1-phosphate and unlike eukaryotic homologs does not act upon the phosphorylated forms of phosphatidylinositol. The main aim of this thesis was to solve the structures of several inactive mutant forms of phospholipase C from Bacillus cereus complexed to myo-inositol which...
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Keresteš, Ondřej. "Biosenzory pro testování inhibitorů cholinesteras." Master's thesis, 2020. http://www.nusl.cz/ntk/nusl-411566.
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Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Student: Bc. Ondřej Keresteš Supervisor: PharmDr. Marta Kučerová, Ph.D. Consultant: pplk. prof. RNDr. Miroslav Pohanka, Ph.D., DSc. Title of diploma's thesis: Biosensors for testing of cholinesterases' inhibitors Acetylcholinesterase (AChE, EC 3.1.1.7) cleaves acetylcholine in the nervous system. Binding of acetylcholine to the receptor and subsequent hydrolytic cleavage at central synapses or neuromuscular junction allows the repeated transmission of nerve impulses. This ensures controlled movement and optimal human cognitive abilities. In case of contact of AChE with the inhibitor, it becomes blocked, thus deteriorating cognitive functions and affects convulsions, with prolonged exposure death can occur. The aim of this work was to test various approaches to measurement of the activity of cholinesterases exposed to two inhibitors, as well as the preparation of biosensors using selected immobilization techniques. Three immobilization procedures were tested. Two of them were using electrochemical detectors and the aim was to optimize the method of quantification activity of AChE by measuring the pH difference (AChE was anchored by the chitosan membrane) and...
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Šobrová, Pavlína. "Diagnostické biosenzory pro encefalopatie způsobené priony." Doctoral thesis, 2013. http://www.nusl.cz/ntk/nusl-176905.
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Prion diseases are fatal transmisible neurodegenerative and infectious disorders (TSEs) of humans and animals, characterized by structural transition of the host-encoded cellular prion protein (PrPC) into the aberrantly folded pathologic isoform PrPSc. The main aim of this work is to summarize present information about prion diseases and their possibilities of determination pointed to electrochemical techniques. For this purpose cyclic voltammetry (CV), differential pulse voltammetry, differential pulse voltammetry Brdicka reaction and chronopotentiometric stripping analysis (CPSA) were used. The estimated detection limits were 32 ug/ml by CV, 16 ug/ml by DPV, 16 ug/ml by DPV -- Brdicka reaction and 8 ug/ml by CPSA. Subsequently, the influence of heat denaturation was observed. It clearly follows from the obtained results that signals of prion decreased linearly depending on the duration of the heat treatment at 99°C for various time intervals 0, 15, 30, 45, and 60 min. Moreover, we aimed our attention on studying of prion protein interaction with CdTe quantum dots (QDs) using electrochemistry. Primarily, we characterized electrochemical properties of QDs and the detection limit at 100 fg/ml was estimated. Further, electrochemical study of prion and QD interactions was carried out to find the most suitable conditions for sensitive detection of prion proteins. Detection limit (3 S/N) was estimated as 1 fg in 5 ul. This makes labeling of proteins with QDs of great importance due to easy applicability and possibility to use in miniaturized devices, which can be used in situ. This should open new possibilities how to determine the presence of these proteins on surgical equipment and other types of materials, which could be contagious.
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Pavlík, Vojtěch. "Návrh a příprava biosenzoru kinázy Src." Master's thesis, 2011. http://www.nusl.cz/ntk/nusl-313045.
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Src kinase is a well-known proto-oncogene that contributes to cell migration and proliferation and is often found deregulated in tumors. Yet, biology of Src is not fully understood and great effort is made to find new tools for broadening knowledge about the kinase. In this thesis is described design and construction of a novel Src biosensor that exploits genetically encodable fluorophores derived from green fluorescent protein and Förster/fluorescence resonance energy transfer (FRET). The fluorophores are inserted directly into the structure of full-length c-Src in the way that should not impair the inner regulatory mechanisms of Src. The created biosensor proved to be sensitive to various stimuli, which also activate c-Src, by increase of activating autophosphorylation on Tyr4162 and decrease in FRET. Preliminary experiments indicate that the Src biosensor can be used to reflect Src activation in fixed cell as well.
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Jabloňků, Jani. "Plasmonické biosenzory v mikro- a nano-škále." Doctoral thesis, 2017. http://www.nusl.cz/ntk/nusl-369544.
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Jurečková, Zuzana. "Detekce oxidačního stresu pomocí elektrochemických DNA biosenzorů." Master's thesis, 2015. http://www.nusl.cz/ntk/nusl-342937.
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Presented Diploma Thesis is focused on the development, characterization, and utilization of simple and inexpensive electrochemical DNA biosensor for the detection of DNA damage caused by oxidative stress. The initial part of the work is devoted to preparation and characterization of a large-surface carbon film electrode (ls-CFE) modified with carbon nanotubes (CNT/ls-CFE). Carbon nanotubes improve electrochemical properties of the transducer and increase the amount of adsorbed DNA on the electrode surface. Testing of the electrode surface modified with multiwalled carbon nanotubes (MWCNT) was performed using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) using a redox system [Fe(CN)6]4-/3- and using square wave voltammetry without any redox indicator. Carbon nanotubes have proved to be unsuitable material for our type of biosensor, but it can be used inanalytical chemistry for the determination of electroactive substances. The second part of this Thesis deals with the application of the prepared DNA biosensor for the detection of DNA damage by oxidative stress. The biosensor based on the ls-CFE was chosen for this purpose, having several advantages, such as its fast preparation, a simple mechanical renewal of the electrode surface, a good reproducibility of measurements,...
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Rodriguez, Emmenegger César. "Senzitivní vrstvy pro optické biosenzory a proteinové čipy." Doctoral thesis, 2012. http://www.nusl.cz/ntk/nusl-307942.
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Sensitive layers for optical biosensors and protein chips The goal of this thesis was the development of sensitive surfaces for optical affinity biosensors detecting in complex biological media. The practical application of these surface-based technologies has been hampered by protein fouling from biological media, in particular blood plasma, where the vast majority of relevant analytes are present. The work of the thesis was centred in three main foci: Design and preparation of antifouling and non-fouling surfaces Evaluation and conceptualisation of their resistance to fouling from blood plasma and serum as well as other biological fluids Preparation of sensitive layers for detection in complex biological media Three approaches were used to prepare protein resistance surfaces, i) ω-functional self-assembled monolayers (SAM), ii) end-tethered polymers and iii) polymer brushes prepared by surface initiated controlled radical polymerisation. Investigation of proteins in the blood plasma deposits on PEG- based surfaces revealed that some fouling is unavoidable in PEG-based surface modifications. A novel type of non-fouling polymer brushes based on poly[N-(2-hydroxypropyl) methacrylamide] challenged the accepted ideas for the design of protein resistant surfaces. For the first time a...
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Drobík, Oto. "Sledování interakce pesticidu s acetylcholinesterasou za použití biosenzoru." Master's thesis, 2011. http://www.nusl.cz/ntk/nusl-297673.
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FOLLOWING OF ACETYLCHOLINESTERASE-PESTICIDE INTERACTION USING BIOSENSOR TECHNOLOGY Oto Drobík Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Organophosphorus and carbamate pesticides are toxic substances used in agriculture as insecticides. These substances are highly toxic for humans as acute poisoning can lead to death via inhibition of acetylcholinesterase. After acetylcholinesterase inhibition, accumulation of acetylcholine follows. In compliance with the thesis aims, functional electrochemical biosensor was constructed. It was based on a standard screen printed amperometric sensor with three electrodes. Herein sensors with platinum and carbon working electrode were tested. Suitable method of electrochemical assay was chosen and some the basic conditions were optimized. Kinetic parameters of enzyme reaction were assessed, too. Various methods of acetylcholinesterase capturing onto sensor were tested and compared one to each other. Paraoxon-methyl and diisopropylfluorophosphate were chosen as the representative of pesticides. Limit of detections for the both pesticides were estimated to be 10-6 - 10-5 mol/l. The prepared biosensors were compared with the standard Ellman's method based on spectrophotometry and the...
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Hemmerová, Erika. "Funkční biomolekulární soubory pro afinitní biosenzory pro medicinální diagnostiku." Doctoral thesis, 2020. http://www.nusl.cz/ntk/nusl-437549.
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OF THE DOCTORAL THESIS Title: Functional biomolecular coatings for affinity biosensors for medical diagnostics Author: Erika Hemmerová Department: Institute of Photonics and Electronics of the Czech Academy of Sciences Chaberská 1014/57, 182 51 Prague, Czech Republic Supervisor: Prof. Ing. Jiří Homola, CSc., DSc. Institute of Photonics and Electronics of the Czech Academy of Sciences Chaberská 1014/57, 182 51 Prague, Czech Republic Abstract: Detection and identification of diseases in their early stages represents one of the major goals of the contemporary medical diagnostics. This need drives a research of biomolecular processes behind the particular diseases and development of analytical devices for routine and long-term monitoring of the individuals' health. Surface plasmon resonance biosensors (SPR) have potential to contribute to addressing both of these challenges. This work aims at advancing multiple aspects of SPR biosensor method. It involves study and optimization of selected functional biomolecular coatings in order to improve the performance characteristics of SPR biosensors. It demonstrates utilization of these coatings in novel detection platforms for sensitive monitoring of multiple analytes, and in research of selected biomolecular interactions related to Alzheimer's disease. The...
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Petržílek, Jan. "Příprava biosenzoru tvorby miRNA efektorového komplexu pomocí CRISPR nukleáz." Master's thesis, 2018. http://www.nusl.cz/ntk/nusl-388379.
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miRNAs are small regulatory RNAs, which function as post-transcriptional mRNA regulators. They direct ribonucleoprotein complexes to cognate mRNA to repress them by translational inhibition and degradation. miRNAs regulate thousands of mRNAs in mammals and have been recognized as regulatory factors in most cellular and developmental processes. Dysregulation of the miRNA pathway can lead to severe defects and diseases. Interestingly, a unique situation exists in mouse oocytes, where all the miRNA pathway components are present, yet the pathway is dispensable and nonfunctional, the molecular foundation of this phenomenon and its significance still remain unclear. In spite of the pronounced effects of the miRNA pathway in gene regulation in somatic cells, study strategies of the pathway bare limitations. Current methods for studying the activity of the miRNA pathway employ corelative studies (such as NGS) or reporter assays, which have relatively low throughput and are prone to artifacts. Here, I present design and development of a new strategy for directly monitor global miRNA pathway activity and integrity in near physiological conditions in living cells, which could also be employed in vivo for studies of mouse oocytes. The strategy is based on fluorescently tagged endogenous proteins of the...
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Šípová, Hana. "Studium biomolekulárních interakcí pomocí optických biosenzorů s povrchovými plasmony." Doctoral thesis, 2014. http://www.nusl.cz/ntk/nusl-338134.
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Surface plasmon resonance (SPR) biosensors represent one of the most advanced sensing technologies for real-time studies of biomolecular interactions. In this thesis, methods for functionalization of SPR substrates were optimized and studied via spectroscopic methods. Effects related to the SPR sensor microfluidic interface on the measured biomolecular interactions were analyzed, and furthermore, means to decrease mass-transport limitations were proposed. Several SPR-based assays regarding the detection of nucleic acids were developed, which allow for the detection of physiologically relevant concentrations of nucleic acids as well as point mutations in a nucleic acid sequence. Assays for the determination of the enzymatic activity of HIV integrase and ribonuclease H were developed. These assays can be employed for the design and synthesis of molecules that function either as antiviral drugs or as gene-regulating agents.
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Navrátil, Jiří. "Vývoj inovativního biosenzoru pro elektrochemickou detekci tumor supresorového genu TP53." Master's thesis, 2019. http://www.nusl.cz/ntk/nusl-427874.
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The theoretical part of the diploma thesis deals with tumour diseases and a general description of tumour suppressor genes. Special attention is paid to the TP53 gene and its influence on the functioning of the organism, including its production of the p53 protein. Based on an analysis and synthesis of findings from specialised sources, the current methods of detecting the aforesaid gene, in particular immunological and elec-trochemical, were described. The last chapter of the theoretical part is devoted to bio-sensors and their classification, with a special focus on electrochemical biosensors. The practical part involves the construction and optimisation of the biosensor, from the cleaning of electrodes, the deposition of gold nanoparticles, the ssDNA bond, the blocking of free binding points and a synthesis of the complementary chain to the resulting measurement and quality testing. This part also presents the concept of an innovative biosensor that offers an easier and more reliable alternative for the detec-tion of the TP53 gene. The last chapter of this part analyses and comments on the aforesaid results.
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Arustamian, Daria. "Jednoduchý elektrochemický DNA biosenzor pro detekci poškození DNA způsobeného UV zářením." Master's thesis, 2018. http://www.nusl.cz/ntk/nusl-380320.
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Ultraviolet (UV) radiation is a common DNA damaging agent. Major DNA lesions, such as cyclobutane pyrimidine dimers (CPDs) and pyrimidine-(6-4)-pyrimidone (6-4PPs) photoproducts, are carcinogenic and mutagenic. UV induced DNA damage was investigated using a simple electrochemical DNA biosensor based on an ultra-trace graphite electrode (UTGE) and low molecular weight doble-stranded DNA (dsDNA) from salmon sperm. Biosensor was prepared using adsorption of dsDNA on a surface of the UTGE and then used to detect UV-induced DNA damage. Effects of UV radiation were investigated using a combination of several electrochemical technics: square-wave voltammetry (SWV) for direct monitoring of DNA base oxidation and cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), as non-direct methods, using redox-active indicator [Fe(CN)6]4-/3- . CV and EIS, which allow characterization of electrode surface, were used to optimize preparation of the dsDNA/UTGE biosensor. Prepared dsDNA/UTGE biosensor was exposed to UV radiation using UV lamp with two set wavelengths: UVC of 254 nm and UVA of 365 nm. UVC radiation was used to damage DNA. Relative signal decrease was 50% after 20 minutes of exposure to UVC radiation. UVA radiation was used to compare effects of different types of UV radiation. Obtained...
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Štipková, Tereza. "Charakteristika vertikálně uspořádaných uhlíkových nanotrubic jako platforem pro aplikace v oblasti biosenzorů." Master's thesis, 2014. http://www.nusl.cz/ntk/nusl-339236.
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Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Tereza Štipková Supervisor: Assoc. Prof. PharmDr. Lucie Nováková, Ph.D. Supervisor specialist: Assist. Prof. Gulnara Safina, Ph.D. Title of thesis: Working performance of vertically aligned carbon nanotubes as platforms for future biosensing applications Blood glucose monitoring is a crucial part of a modern diabetes therapy. In this work, the multi-walled vertically aligned on-chip grown carbon nanotubes (CNT) were applied in construction of a novel third-generation biosensor. The main focus of this work was to investigate the possibility of future application of the sensor in glucose monitoring. The CNT- based amperometric biosensor was developed by immobilising cellobiose dehydrogenase from Corynascus thermophilus onto the CNT surface by physical adsorption. The highly hydrophobic surface of the nanotubes was functionalized with 0.0001% (w/v) polyvinyl alcohol (PVA) in order to enable the enzyme adsorption. X-ray photoelectron spectroscopy measurements confirmed the successful enzyme immobilization onto PVA modified CNT surface. The surface topography and elemental composition of the nanomaterial were examined by scanning electron microscopy and energy-dispersive X-ray spectroscopy...
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Stávková, Klára. "Využití elektrochemického DNA biosenzoru při detekci poškození DNA způsobeného genotoxickým 2-nitrofluorenem." Master's thesis, 2014. http://www.nusl.cz/ntk/nusl-340403.
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2-Nitrofluorene is a model representative of nitrated polycyclic aromatic hydrocarbons (NPAH) which belongs to a group of mutagens and carcinogens. Interaction of DNA with genotoxic 2-nitrofluorene was monitored by an electrochemical DNA biosensor made of a glassy carbon electrode (GCE) and low molecular weight DNA from salmon sperm. Techniques used are electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and square-wave voltammetry (SWV). Using the EIS technique, no damage to DNA, which would cause strand breaks in DNA, was observed, whereas using the CV technique, the intercalation of NF to the structure of DNA was observed, leading to the formation of a NF-DNA complex. The intercalation results in a reduction of electroactive sites which can be oxidized. It was verified using the SWV technique, by which a decrease of the peak heights of adenosine and guanosine was observed. Because of the dangerous effect of NF on the structure of DNA, an electroanalytical method for its determination was developed. An applicability of the method was successfully tested on a model sample of sand. For the development of the technique, differential pulse voltammetry (DPV) was used in a mixture of the Britton-Robinson buffer of pH 7.0 and ethanol in a ratio of 7:3 (v/v) and with a periodic...
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Blašková, Marta. "Využití jednoduchého elektrochemického DNA biosenzoru při stanovení environmentálních polutantů a vyšetřování jejich interakce s DNA." Master's thesis, 2014. http://www.nusl.cz/ntk/nusl-333185.
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The interaction between three selected representatives of environmental pollutants - naphthalene, anthracene, and 2-aminoanthracene - and DNA was investigated using an electrochemical DNA biosensor based on a glassy carbon electrode (GCE) and low molecular weight DNA from salmon sperm (DNA/GCE). The interactions with DNA were monitored using square wave voltammetry (SWV) and electrochemical impedance spectroscopy (EIS). For naphthalene, there was no DNA damaging interaction observed. In the case of anthracene, the formation of an intercalation complex [DNA-anthracene] was observed. However, its formation does not cause DNA strand breaks. The formation of similar intercalation complex was observed for 2-aminoanthracene [DNA-2-aminoanthracene], where we suppose on the basis of the results obtained that the intercalation of 2-aminoanthracene into the DNA double helix induces a tension and subsequent formation of single-strand breaks, which cause that the fragments of DNA fall away from the electrode surface. The intercalative interaction of DNA with anthracene a 2-aminoanthracene was used in the development of electrochemical methods for determination of these compounds at the GCE and DNA/GCE. At the development of the methods, DC voltammetry (DCV) and differential pulse voltammetry (DPV) were used....
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Bocková, Markéta. "Multifunkční biomolekulární soubory pro paralelizovanou analýzu biomolekulárních interakcí." Doctoral thesis, 2019. http://www.nusl.cz/ntk/nusl-404427.
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Title: Multifunctional biomolecular assemblies for parallelized analysis of biomolecular interactions Author: Markéta Bocková Department / Institute: Institute of Physics, Charles University Supervisor of the doctoral thesis: Prof. Jiří Homola, Ph.D., DSc., Institute of Photonics and Electronics, The Czech Academy of Sciences Abstract: Surface plasmon resonance (SPR) biosensors represent the most advanced optical method for the direct, real-time monitoring of biomolecular interactions without the need for labelling. This doctoral thesis aims to advance the SPR biosensor method and to expand its utility in the investigation of biomolecular interactions. This encompasses activities on two major fronts of SPR biosensor research - immobilization methods and biosensing methodologies. Methods for the immobilization of biomolecules were researched with the aim of enabling the immobilization of a broad range of biomolecules on the SPR biosensor surface in a spatially controlled manner. The development of novel biosensing methodologies was pursued in order to address the current limitations of SPR biosensors associated with non-specific adsorption and limited analyte transport, and thus to improve the accuracy and robustness of SPR biosensor measurements. Finally, advances in the development of immobilization...
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Vopálenská, Irena. "Diferenciace kolonií kvasinek a vývoj nových přístupů pro monitorování dostupnosti kyslíku a přítomnosti živin." Doctoral thesis, 2015. http://www.nusl.cz/ntk/nusl-266322.
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Yeast Saccharomyces cerevisiae as an unicellular organism is one of the best-studied experimental organisms. It is an important model organism for the study of intracellular processes of eukaryotic cells. Yeasts are also social organisms with cell-to-cell communication able to form organized multicellular structures (colonies and biofilms). Yeast and other microorganisms in nature prefer to form colonies on solid substrates rather than to grow as "planktonic" single cells (Palková, 2004; Wimpenny, 2009). The yeast S. cerevisiae typically forms colonies, biofilms were described only rarely. Yeast colonies exhibit an organized morphological pattern characteristic of each particular yeast strain (Kocková-Kratochvílová, 1982). This work is focusing on morphology and differentiation of the S. cerevisiae colonies of common laboratory strains forming less structured colonies, and strains of the Σ1278b genetic background forming highly structured "fluffy" colonies. It shows that polarized budding pattern and especially cell ability to form aggregates enable development of structured morphology. During development of "fluffy" colonies two differently regulated events of dimorphic switch from yeast form to filamentous growth occur. One of these events is dependent on the surface glycoprotein, Flo11p flocculin. This...
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Hájková, Andrea. "Nové přístupy při elektrochemickém stanovení cizorodých látek a studiu jejich interakce s DNA." Doctoral thesis, 2016. http://www.nusl.cz/ntk/nusl-352074.
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Presented Ph.D. Thesis is focused on the development of analytical methods applicable for determination of selected xenobiotic compounds and for monitoring DNA damage they can induce. The main attention has been paid to the development and testing of non-toxic electrode materials for preparation of miniaturized electrochemical devices and novel electrochemical DNA biosensors. 2-Aminofluoren-9-one (2-AFN) was selected as a model environmental pollutant, which belongs to the group of hazardous genotoxic substances. Its carcinogenic and mutagenic effects may represent a risk to living and working environment. 2-AFN has one oxo group, where the cathodic reduction occurs, and one amino group, where the anodic oxidation occurs. The voltammetric behavior of 2-AFN in the negative potential region was investigated at a mercury meniscus modified silver solid amalgam electrode (m-AgSAE) representing a non-toxic and more mechanically robust alternative to mercury electrodes. This working electrode was subsequently used for the development of a newly designed miniaturized electrode system (MES), which has many benefits as the possibility of simple field measurements, easy portability, and the measurement in sample volume 100 µL. Moreover, a glassy carbon electrode (GCE) was used for further investigation of...