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4 years ago

Surface-enhanced Raman scattering detection of Amyloid-β using a ...

Surface-enhanced Raman scattering detection of Amyloid-β using a ...

Abstract In this Master

Abstract In this Master thesis, amyloid-β (Aβ) oligomers, which are considered as the main pathogenic agents in the Alzheimer’s disease, were trapped using a nanogap enabled dielectrophoretic device and detected with surface-enhanced Raman spectroscopy. Fluorescence observation of the accumulation of thioflavin T (ThT)-labeled Aβ oligomers and FAM-labeled Aβ oligomers at the nanogap between electrodes provided evidence of the trapping. Additionally, the trapping event was monitored with nanoelectronic current measurements confirming the presence of the molecules in the vicinity of the gap. Surface-enhanced Raman spectra of ThT-labeled Aβ oligomers were taken and analyzed. Finally, two fabrication methods that incorporate a microfluidic channel of 7 mm length, 250 m width and 50/70 m depth to the nanogap device have been developed, one using PDMS and the other using double-sided tape as material; which potentially enable next generation experiments with fluidic exchanges, applied transversal biases etc. Kurzzusammenfassung Im Rahmen dieser Masterarbeit werden Amyloid-β (Aβ) Oligomere, welche als Haupterreger für die Alzheimer-Krankheit angesehen werden, mithilfe eines Biosensors mittels Dielektrophorese in einem „Nanogap“ eingefangen und mit oberflächenverstärker Ramanspektroskopie detektiert. Fluoreszenzbeobachtungen der Akkumulation von thioflavin T (ThT)-markierten Aβ Oligomeren und FAM-markierten Aβ Oligomeren am Nanogap zwischen zwei Elektroden dienten als Nachweis für das erfolgreiche Einfangen derselben. Diese Ereignisse wurden zudem mittels nanoelektronischen Strommessungen überprüft, und die Gegenwart von Molekülen nahe dem Nanogap konnte nachgewiesen werden. Oberflächenverstärkte Ramanspektren von ThT-markierten Aβ Oligomeren wurde aufgenommen und analysiert. Schließlich wurden zwei Herstellungsmethoden entwickelt, die einen mikrofluidischen Kanal mit einer Länge von 7 mm, einer Breite von 250 µm und einer Tiefe von ca. 50 bzw. 70 µm in das Nanogap-Bauteil integrieren, wobei in der einen Variante PDMS und in der anderen doppelseitiges Klebeband als Material verwendet wird. Dies ermöglicht zukünftige Experimente mit Flüssigkeitsaustausch, angelegten transversalen Vorspannungen usw. i

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