AFM-STED correlative nanoscopy provides a new view on the formation process of misfolded protein aggregates

The main part of my PhD work focused on the application of an advanced integrated technique, based on the coupling of an atomic force microscope (AFM) and a stimulated emission depletion (STED) microscope in the study of amyloid fibrils formation. This coupled system allows the acquisition of super-resolution fluorescence images, perfectly overlapped with AFM topography. Exploiting the extended capability offered by this technique, I highlighted some important features on the mechanisms followed by the labeled and unlabeled proteins through their aggregation pathway. The results demonstrates that labeled molecules are involved only in selected pathways of aggregation, among the multiple that are present in the aggregation reaction. In a second part of my work, I investigated the process of interaction between Alpha-synuclein (α-Syn), the pathological peptide associated to the Parkinson’s disease, and model lipid membranes. The aim of this study was to identify molecular mechanisms that are indicated as the base of neurodegeneration, not only in Parkinson’s disease, but also in a large class of disorders, indicated as protein misfolding diseases.