Finalist EPFL doctorate Award 2014 - Paolo Annibale
Special distinction from the selection committee to Paolo Annibale for his thesis "Investigating the Impact of Single Molecule Fluorescence Dynamics on Photo Activated Localization Microscopy Experiments". Thesis n° 5517 (2012). Thesis director: Prof. A. Radenovic
When fluorophores are densely packed in a biological sample, localizing them one at a time is one of the paths to optical super-resolution.
Photo Activated Localization Microscopy (PALM) is one of the methods of this burgeoning field that presents a large promise for addressing physical and biological questions requiring non-invasive, high-resolution optical imaging at the nanoscale.
The principal theme of this thesis is the investigation of the single molecule fluorescence dynamics of the fluorophores used. Beginning with a discussion of the properties of a bright fluorescent protein, mEos2, we propose an original approach based on the use of the temporal information besides the spatial one when treating PALM data. This allows both a more accurate quantification of the number of fluorophores activated in the sample as well as the correct identification of relevant biological structures, such as clusters, on the plasma membrane of cells.
The second theme is the application of PALM to probe the functional arrangement of an important class of cell membrane proteins, through the study of the prototypical G protein-coupled receptor beta2-Adrenergic Receptor. PALM is then shown to have the potential to be included in the palette of the available techniques to study membrane protein organization.
