MultiSmartSTED – Multiscale smart stimulated emission depletion microscopy

• Applicant: Simone Vera Ezendam
• Subject Area: Biophysics ; Optics, Quantum Optics and Physics of Atoms, Molecules and Plasmas
• Term: since 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 549029507

Cells are incredibly complex machinaries with processes occuring at different speeds and across various lengthscales, from nanosecond to seconds and from nanometers to micrometers. However, our current methods do not allow us to observe these dynamics comprehensively in real-time. This project seeks to overcome this limitation by introducing a novel technique named STARSS STED, which will be integrated into a sophisticated microscopy framework referred to as MultiSmartSTED (Multiscale Smart STED Microscopy). The primary objective is to develop STARSS STED, inspired by recent advancements in fluorescence microscopy, notably recognized with the Nobel Prize for super-resolution microscopy in 2014. These techniques enable imaging at the nanometer scale, with STED (Stimulated Emission Depletion) microscopy being one prominent example. STARSS STED innovatively utilizes simulated emission to enhance fluorescence anisotropy measurements. This approach allows for the study of rotational diffusion - a rapid process offering insights into cellular membrane dynamics and composition. Consequently, STARSS STED promises to unveil new insights into fundamental biological processes at the molecular level. The secondary aim involves seamlessly integrating STARSS STED into the MultiSmartSTED framework. By leveraging advanced algorithms and automation, MultiSmartSTED optimizes imaging parameters on-the-fly, enables real-time adaptability and consistent monitoring of cellular regions. This integration will bring together three modalities: widefield microscopy, STED microscopy, and STARSS STED. Together, these modalities offer unprecedented capabilities to explore a wide range of spatio-temporal scales, enabling researchers to gain deeper insights into the dynamic behaviors of living cells.

• DFG Programme: WBP Fellowship
• International Connection: Sweden
• Host: Professorin Dr. Ilaria Testa