Chování hydrofilní fluorescenční sondy Oregon Green 488 v prostředí hydrogelu
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Vysoké učení technické v Brně. Fakulta chemická
Abstract
Táto bakalárska práca sa zaoberá štúdiom chovaní hydrofilnej fluorescenčnej sondy Oregon Green 488 v prostredie hydrogélov. Hydrogél bol vytvorený použitím hyaluronanu sodného a CTAB (cetyltrimethylammonium bromid), ktorý je kationtový tenzid. Ako rozpúšťadlo bol použitý fyziologický roztok NaCl s koncentráciou 0,15 M. Pre charakterizáciu chovaní fluorescenčnej sondy bol použitá technika stacionárnej fluorescencie a časovo rozlíšenej fluorescencie. Metódou TRES (časovo rozlíšené emisné spektrá) boli preskúmané emisné spektra fluorescenčnej sondy a nájdené emisné maxima. Pomocou parametra poklesu intenzity fluorescencie pri daných vlnových dĺžkach bola vytvorená dekonvolúcia emisného spektra, ktorá poskytovala informácie o dobe života a relatívnom zastúpení jednotlivých stavov fluorescenčnej sondy v roztoku. Hydrogély boli skúmané 5 dní a 1 mesiac po ich vytvorení a získané výsledky boli porovnané. Oregon Green 488 vykazoval jednu dobu života v prostredie hydrogélov, ktorá sa pohybovala v rozmedzí 4,1–4,45 ns v čerstvom géle, a 4,14,6 ns v stálom géle. Emisné maximum bolo nájdené v bode 525 nm. Fluorescenčná sonda sa chová v hydrogéloch ustálene a stabilne, pravdepodobne nedochádza k reakcii medzi sondou a gélom. Nebola zaznamenaná interakcia medzi hyaluronanom a sondou.
The aim of this bachelor’s thesis is to study the behavior of a hydrophilic fluorescent probe named Oregon Green 488 in hydrogel environment. The hydrogel was created using sodium hyaluronan and CTAB (cetyl trimethylammonium bromide) as a cationic surfactant. Solution of 0.15 M NaCl was used as a solvent, which provides physiologic conditions in the sample. The behavior of the fluorescent probe was characterized by steady-state and time-resolved fluorescent measurements. To obtain the emission spectra of the probe and to find the emission maximum the method named TRES (time-resolved emission spectra) was used. Fluorescence intensity decay measurements as a function of wavelengths allowed to create deconvolution of the emission spectra, which provided information about the fluorescent lifetime and the probe’s states’ relative representation in the solution. Hydrogels were examined 5 days and 1 month after the preparation and the results were compared. Oregon Green 488 demonstrated one fluorescent lifetime in the hydrogel environment in the range, 4,14,45 nm in the fresh hydrogel and 4,14,6 nm in the stable hydrogel. The emission maximum was obtained at 525 nm in both cases. The fluorescent probe’s behavior in the hydrogel environment is stable, presumably there are no interactions occurring between the hydrogel and the probe. Furthermore, no interactions were recorded between the probe and hyaluronan.
The aim of this bachelor’s thesis is to study the behavior of a hydrophilic fluorescent probe named Oregon Green 488 in hydrogel environment. The hydrogel was created using sodium hyaluronan and CTAB (cetyl trimethylammonium bromide) as a cationic surfactant. Solution of 0.15 M NaCl was used as a solvent, which provides physiologic conditions in the sample. The behavior of the fluorescent probe was characterized by steady-state and time-resolved fluorescent measurements. To obtain the emission spectra of the probe and to find the emission maximum the method named TRES (time-resolved emission spectra) was used. Fluorescence intensity decay measurements as a function of wavelengths allowed to create deconvolution of the emission spectra, which provided information about the fluorescent lifetime and the probe’s states’ relative representation in the solution. Hydrogels were examined 5 days and 1 month after the preparation and the results were compared. Oregon Green 488 demonstrated one fluorescent lifetime in the hydrogel environment in the range, 4,14,45 nm in the fresh hydrogel and 4,14,6 nm in the stable hydrogel. The emission maximum was obtained at 525 nm in both cases. The fluorescent probe’s behavior in the hydrogel environment is stable, presumably there are no interactions occurring between the hydrogel and the probe. Furthermore, no interactions were recorded between the probe and hyaluronan.
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Citation
LACKO, K. Chování hydrofilní fluorescenční sondy Oregon Green 488 v prostředí hydrogelu [online]. Brno: Vysoké učení technické v Brně. Fakulta chemická. 2015.
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Document version
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cs
Study field
Chemie pro medicínské aplikace
Comittee
prof. Ing. Miloslav Pekař, CSc. (předseda)
prof. RNDr. Ivana Márová, CSc. (místopředseda)
prof. Ing. Oldřich Zmeškal, CSc. (člen)
prof. Ing. Michal Veselý, CSc. (člen)
doc. Ing. Pavel Diviš, Ph.D. (člen)
Date of acceptance
2015-06-19
Defence
Pekař – vliv micel na spektra
Diviš – charakter spekter AES
Result of defence
práce byla úspěšně obhájena
Document licence
Standardní licenční smlouva - přístup k plnému textu bez omezení