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Analysis of Thick Film Amperometrical Sensors Signal and Its Usage for Measurement and Characterization of Enzymes
Ondruch, Vít ; Kizek, René (referee) ; Masojídek,, Jiří (referee) ; Vrba, Radimír (advisor)
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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Biosensors based on functionalized graphene
Pavlásková, Lucie ; Skládal, Petr (referee) ; Bartošík, Miroslav (advisor)
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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Microelectrode arrays for mioelectronic
Bráblíková, Aneta ; Vala, Martin (referee) ; Salyk, Ota (advisor)
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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DESIGN OF FIBER-OPTIC BIOSENSOR WITH NEAR-INFRARED SPECTRAL ANALYSIS
Křepelka, Pavel ; Jakubec,, Martin (referee) ; Skládal, Petr (referee) ; Mikulka, Jan (advisor)
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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Optimalization of printing methods of organic semiconducting layers preparation
Ehlich, Jiří ; Vala, Martin (referee) ; Salyk, Ota (advisor)
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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Reliability analysis of substance measurements using biosensors - limit of detection robust method design
Láznička, Jan ; Brno, Karel Lacina, MU (referee) ; Grosmanová, Zuzana (advisor)
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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Printed Biosensor Based on Organic Electrochemical Transistor
Omasta, Lukáš ; Mikula, Milan (referee) ; Boušek, Jaroslav (referee) ; Salyk, Ota (advisor)
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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Technology of wearable sensors
Javor, Adrián ; Majzlíková, Petra (referee) ; Hubálek, Jaromír (advisor)
This bachelor thesis deals with thick-film and thin-film technology of creating wearable sensors on human body, that sense substances in perspiration. Part is devoted to their classification and making subsequent proposal of flexible three-electrode biosensors using 4 different electrochemical transducers. All of these biosensors sense concentration of lactate in human sweat, and are based on immobilization lactate oxidase enzyme on the working electrode. Technological process of production is based on combination of thick-film and thin-film technology. Conclusion of this bachelor thesis contains amperometric evaluations of the lactate concentration, from which calibration curves were created.
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Application of metal and semiconductor nanostructures for biodetection
Kejík, Lukáš ; Skládal, Petr (referee) ; Kolíbal, Miroslav (advisor)
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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