Please use this identifier to cite or link to this item:https://hdl.handle.net/20.500.12259/107567
Type of publication: bachelor thesis
Field of Science: Chemijos inžinerija / Chemical engineering (T005)
Author(s): Liudžiūtė, Gintarė
Supervisor: Rodaitė - Riševičienė, Raminta,
Title: Aukštosios įtampos impulsų sukelti efektai ir jų pasekmės fluorescuojančiam vandenilio peroksido indikatoriui Amplekso raudonojam bei terpės pH
Other Title: Effects and results caused by high voltage impulses on Amplex red – fluorogenic probe of hydrogen peroxide, and pH of solution
Extent: 36 p.
Date: 18-Jun-2020
Keywords: Elektroporacija;Electroporation;Amplekso raudonasis;Amplex red;Membrana;Membrane
Abstract: Grįžtamosios elektroporacijos metu ląstelės membranoje yra sukuriamos poros, per kurias į ląstelės vidų gali būti įvedami įvairūs vaistai ar kitos medžiagos. Tačiau šis metodas turi neigiamų šalutinių efektų. Vienas jų, kuomet elektroporacijos metu iš elektrodų į terpę išeinantys metalų jonai, pavyzdžiui Fe2+ ir Fe3+, gali pastebimai pakeisti elektroporuojamų terpių pH. Siekiant atkartoti šį procesą buvo pasitelkti vandeniniai FeCl2 ir FeCl3 druskų tirpalai ir įsitikinta, kad geležies jonai mažina buferinių bei elektroporacijoje naudojamų terpių pH. Taip pat elektrinių impulsų metu gali susidaryti aktyviosios deguonies formos, kitaip ADF, kurioms priskiriamas ir vandenilio peroksidas. Todėl prieš nustatant kokias pasekmes jis sukelia ląstelėms, būtina jį identifikuoti. Tai padaryti galima naudojant fluorescuojantį indikatorių – Amplekso raudonąjį. Šio indikatoriaus pagalba vandenilio peroksidas gali būti ne tik atpažintas, bet ir nustatytas jo kiekis. Pagrindinis tyrimų tikslas buvo ištirti aukštosios įtampos impulsų sukeltų efektų pasekmes fluorescuojančiajam vandenilio peroksido indikatoriui Amplekso raudonajam ir terpės pH. Nustatėme, kad po pritaikytų 8 stačiakampių, 100 µs trukmės, 0,2 – 1,4 kv/cm elektrinių impulsų, proporcingai didėjo Amplekso raudonojo fluorescencijos intensyvumas, dėl PBS terpėje susidariusio vandenilio peroksido. / During reversible electroporation, pores are created in the cell membrane due to an applied electric field. This membrane permeabilization could help introduce various drugs or other substances into the cell. However, this method has negative effects on cell viability. One of them is when metal ions, such as Fe2 + and Fe3 +, solubilize from the electrodes into the medium. It can significantly change the pH of the electroporated media. To repeat this process were used aqueous solution FeCl2 and FeCl3. The experiment results confirmed that iron ions lower the pH of the buffer and electroporation media. Also, electrical pulses can form reactive oxygen species, in other words, ROS, which include hydrogen peroxide. Therefore, before further research on side effects for cells of hydrogen peroxide, it is a must to identify it. This can be done using a fluorogenic probe - Amplex Red. Using this indicator, hydrogen peroxide can not only be identified but also determined. The main object of the research was to determine the effect of electroporation on the fluorescence of Amplex red. For this purpose, an experiment was carried out, during which it was ascertained that by using 8 100 microseconds duration of increasing high pulses electric field. The results show that increasing the electrical voltage also increases the fluorescence of Amplex red, which means that the concentration of hydrogen peroxide is getting bigger.
During reversible electroporation, pores are created in the cell membrane due to an applied electric field. This membrane permeabilization could help introduce various drugs or other substances into the cell. However, this method has negative effects on cell viability. One of them is when metal ions, such as Fe2 + and Fe3 +, solubilize from the electrodes into the medium. It can significantly change the pH of the electroporated media. To repeat this process were used aqueous solution FeCl2 and FeCl3. The experiment results confirmed that iron ions lower the pH of the buffer and electroporation media. Also, electrical pulses can form reactive oxygen species, in other words, ROS, which include hydrogen peroxide. Therefore, before further research on side effects for cells of hydrogen peroxide, it is necessary to identify it. This can be done using a fluorogenic probe - Amplex Red. Using this indicator, hydrogen peroxide can not only be identified but also determined. The main object of the research was to determine the effect of electroporation on the fluorescence of Amplex red. For this purpose, an experiment was carried out, during which it was ascertained that by using 8 100 microseconds duration of increasing high pulses electric field. The results show that increasing the electrical voltage also increases the fluorescence of Amplex red, which means that the concentration of hydrogen peroxide is getting bigger.
Internet: https://hdl.handle.net/20.500.12259/107567
Appears in Collections:2020 m. (GMF bak.)

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