Assessing cytoskeletal network architecture and co-localization with microscopy (MEP)
The cytoskeleton of the cell confers the cell with shape and strength to hold, as well as the ability to deform. The balance between strength and deformability arises from the interaction between the different cytoskeletal families (actin, microtubules, intermediate filaments). We refer to the interplay between the different filaments as cytoskeletal crosstalk. Crosstalk is not only mediated by direct or indirect interactions between the filaments, but also by some proteins that “bond” them together, which we call crosslinkers. In our lab, we have engineered a library of crosslinkers in order to test and see what properties emerge from the interaction between different cytoskeletal filaments.
In this project you will use Total Internal Reflection Fluorescence microscopy (TIRF) and Transmission Electron Microscopy (TEM) to study the interactions emerging from the different cytoskeletal filaments by checking with microscopy the co-localization of the different cytoskeletal filaments, as well as the architecture of the network under different conditions of crosslinking and filament composition (which will affect the persistence length, or flexibility). In this project you will learn how to work with in vitro reconstituted cytoskeletal proteins, use the TIRF and TEM microscopes and analyse and interpret your data.