Innovative fluorescence light microscopy techniques including quantitative live cell imaging and super resolution (SR) microscopy, greatly improve our ability to visualize protein complexes at high spatial and temporal resolution in their intact environment. Such information is critical for understanding how proteins mediate their function in the complex and packed environment of the cell.

We adapt and tailor state-of-the-art microscopy techniques to study the mechanism of molecular machines that drive membrane fission in cells. Specifically, we focus on the ESCRT membrane fission machine which is known to mediating membrane remodeling in a variety of processes in cells including biogenesis of multivesicular bodies, viral budding, cell division and membrane repair. Interestingly, the ESCRT complex is composed of over 20 different proteins that interact with one another but how they organize in cells to orchestrate the energetically unfavorable reaction of membrane fission in cells is unknown.

While much information has been obtained in recent years on the ESCRTs using high resolution imaging techniques, a mechanical understanding of membrane fission in a physiological context is still missing, a shortcoming that stresses the need to increase the spatiotemporal resolution and improve the live cell capabilities of current imaging techniques. By constantly exploring new microscopy techniques and by tailoring current techniques to our specific questions we aim to obtain the information needed for understanding how ESCRTs operate in their native, physiological environment.