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Označevanje polielektrolitov s fluorescentnimi barvili in spremljanje elektrostatskega oblaganja bakterijskih celic z mikroskopijo in kalorimetrijo = Labelling of polyelectrolytes with fluorescent dyes and monitoring of electrostatic coating of bacterial cells with microscopy and calorimetry : enoviti magistrski študij farmacijeGrilc, Nina KatarinaŽive bakterijske celice so danes nepogrešljive na mnogih področjih biotehnologije in biomedicine. Za te namene pogosto spreminjamo površinske lastnosti celic. Ena od možnosti za modifikacijo celične ... površine je oblaganje celic z metodo nanoplastenja, pri kateri se plasti polielektrolitov nalagajo na nabito površino oblaganca na podlagi elektrostatskih interakcij. V magistrski nalogi smo spremljali tvorbo polikationske plasti na negativno nabiti površini bakterijskih celic. To je prva naloga, ki preučuje nanoplastenje celic s fluorescenčno mikroskopijo z vzbujenim praznjenjem emisije (STED). Namen je bil optimizacija postopka označevanja polikationov s fluorescentnim barvilom in izbor barvila, ki na uporabljenem STED mikroskopu izkazuje boljšo ločljivost metode kot na konfokalnem mikroskopu. Sistematično smo nanoplastenje s polielektroliti preučili na dveh vrstah celic (E. coli in P. stutzeri) z meritvijo zeta potenciala (ZP) in STED mikroskopijo. Nanooblaganje smo spremljali še z izotermno titracijsko kalorimetrijo (ITC). Rezultati kažejo izboljšanje ločljivosti na STED v primerjavi s konfokalnim mikroskopom pri barvilu STAR RED, ne pa pri ATTO 594 in rodamin-B-izotiocianatu. S STAR RED smo označili polikatione (visoko- in nizkomolekularna polietilenimin - PEI in polialilamin klorid - PAH) in z njimi obložili celice E. coli in P. stutzeri. ZP celic se je spremenil iz negativnega pred nanooblaganjem v pozitivnega po nanooblaganju. STED slike kažejo nastanek nanoobloge na celični površini. Ugotovili smo, da polikationi prehajajo v notranjost P. stutzeri, ne pa v E. coli, nismo pa dokazali razlik v prehajanju polikationov glede na molekulsko maso. Pri ITC so krivulje odvisnosti toplotnega toka od količine dodanega polielektrolita pri E. coli dosegale plato, pri katerem predpostavimo zasedenost celotne celične površine s polikationom. Pri P. stutzeri je bilo prisotno sipanje podatkov okoli pričakovane (sigmoidne) oblike krivulje zaradi prehajanja polielektrolita v notranjost. Nanooblaganje preučevanih bakterijskih celic s PEI in PAH je bilo uspešno, saj se je površinski naboj spremenil iz negativnega v pozitivnega, tvorba nanoobloge pa je vidna tudi s STED. Permeabilnost celične stene za polikation je manj odvisna od njegove molekulske mase in bolj od vrste celic, kar smo dokazali z opazovanjem lokalizacije polikationov po nanooblaganju s STED mikroskopijo. Z ITC potrdimo doseganje nasičenja celične površine s polikationom.Type of material - master's thesis ; adult, seriousPublication and manufacture - Ljubljana : [N. K. Grilc], 2020Language - slovenianCOBISS.SI-ID - 4878705
Link(s):
https://repozitorij.uni-lj.si/IzpisGradiva.php?id=113691Vsebina dokumenta nedostopna do 24.01.2021
Author
Grilc, Nina Katarina
Other authors
Kristl, Julijana |
Mirtič, Janja, farmacevtka
Topics
Bakterije |
Mikroskopiranje |
Nanomateriali |
nanooblaganje |
polielektrolit |
STED |
fluorescenčna mikroskopija z vzbujenim praznenjem emisije |
kalorimetrija
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Library | Call number – location, accession no. ... | Copy status |
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Faculty of Pharmacy, Lj. | Knjižnica emšf 66.02 GRILC N. K. Označevanje 813 IN: 0016463 |
available - reading room |
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Grilc, Nina Katarina | ![]() |
Kristl, Julijana | 09032 |
Mirtič, Janja, farmacevtka | 39199 |
Source: Personal bibliographies
and: SICRIS
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