Partenaires
lundi 20 janvier
Mise à jour
mercredi 24 juillet 2024
Accueil > Production Scientifique > Thèses soutenues > Année 2024 > Soutenance de thèse de Lancelot Pincet (31 mai)
Soutenance de thèse de Lancelot Pincet (31 mai)
par - 28 mai 2024 (modifié le 24 juillet 2024)
Dynamic excitation systems for quantitative and super-resolved fluorescence microscopy.
Single Molecule Localization Microscopy (SMLM) is a super-resolution optical technique enabling the observation of biological samples labeled with fluorescent dyes at resolutions well below the diffraction limit. The quality of this imaging heavily relies on the ability to observe molecules individually, requiring precise control of fluorescent dye photophysics for them to emit with a high sparsity in both space and time. Until now, dynamic excitation methods aimed to produce uniform illumination over large fields (200x200 um²). However, these types of illumination encounter difficulties in imaging dense biological samples, such as neurons, where the diversity in dye density prevented the generation of a uniform single molecule regime across the entire observed area. To address this issue, I propose a new approach that dynamically adjusts illumination based on sample density. This method combines a novel tri-dynamic optical excitation system with a feedback loop based on density analysis, benefiting from an in-depth study of fluorescent dye photophysics. The intelligent imaging system, where the excitation pattern varies over time, integrates a 2D scanning system, a variable zoom system, and a laser. This allows for the generation of a variety of dynamically changing illumination patterns to adapt to the observed sample and the density of locally detected localizations. This new approach has been validated on various biological samples. Additionally, the dynamic excitation system has also been explored for live samples imaging techniques, such as MSIM or FRAP.