Super-resolution flourescence microscopy
The pre-synaptic protein α-synuclein (α-syn) can be found in amyloid plaques in patients with Parkinson’s Disease (PD). However, the mechanisms behind how α-syn is involved in the course of PD is still unknown. An important event seems to be the interaction between a-syn and lipids, which are the main building block of the membrane surrounding each cell. The aim of my project is to use fluorescence microscopy to obtain information about possible vesicle fusion and exchange of lipids between single, fluorescently labelled lipid vesicles, both due to interactions with -syn. This is done using vesicles containing lipids labelled with two different dyes forming a fluorescence resonance energy transfer (fret) pair. Lipid exchange or fusion between two vesicles labeled with different dyes can be detected by colocalization of the dyes and the appearance of a fret signal when exciting the donor dye. It is possible to distinguish between fusion and exchange since a fusion event would lead to fewer but larger vesicles with higher fluorescence intensities, while an exchange event would lead to an unchanged number of vesicles with lower fluorescence intensities of respective dye.