Kurilchenko I. Fluorescent labeling of nano-scale carriers for tracing their interaction with biological objects.

The thesis is devoted to the development of the method of fluorescent labeling of nano-scale carriers (surfactant liposomes and micelles) for long-time tracing of the nanocarriers interaction with living cells in vitro experiments. The method is based on the effect of Foerster Resonance Energy Transfer (FRET) between several fluorescent probes preloaded in the nanocarrier and -ratiometric detection of energy donor and acceptor fluorescence signals using microspectoscopy technique. The interaction of fluorescent probes (polymethine and squaraine dyes) interaction in a nanovolume of phosphatidylcholine (PC) liposomes and sodium dodecylsulfate (SDS) micelles has been studied in detail. It has been found that the ability of the fluorescent probes to interact with each other in the nanovolume should be taken into consideration for creation of labeling FRET-pairs. The evaluation methods for fluorescent probes accumulation in cells using total fluorescent image brightness has been developed. It has been shown that liposome FRET-labeling allows the differences in dynamics and efficiency of the nanocarriers interaction with cells to be detected, which associated with the different composition of cell plasma membranes for different cells, their different viscosity, cell metabolic and functional activity.