Lokalizace fluorescenčních značek a určování velikostí lipidových nanodomén pomocí moderních fluorescenčních metod

Šachl, Radek

iii Organizations Umeå University Department of Chemistry, Umeå, SE & Charles University in Prague, Faculty of Science, Prague, CZ Document name Doctoral thesis Date of issue February 2012 Author Radek Šachl Title Localisation of Fluorescent Probes and the Estimation of Lipid Nanodomain Sizes by Modern Fluorescence Techniques Abstract The thesis is divided into two major parts. The first part focuses on the localisation of probes in lipid/polymeric bilayers and in GM1 micelles. Included in this thesis is a new approach based on electronic energy transfer/migration (FRET/DDEM), which efficiently determines transversal positions of fluorescent molecules in lipid bilayers. This approach has been used to locate newly synthesized lipid probes in DOPC bilayers. The label was introduced at the end of sn-2 acyl chains of variable length. Analytical models accounting for FRET exist for a limited number of basic geometries. Here, a combination of FRET and Monte Carlo simulations enables the localisation of probes in bicelles and in bilayers containing pores, i.e. in lipid systems with variable curvature, or in non-homogenous lipid systems. This approach has been used to test whether conical-like fluorescence probes have an increased affinity to highly curved regions, which would enable preferential labelling of...

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