Midterm colloquium Koen Stapel

Optimal illumination pattern in iterative modulation enhanced single molecule localization

Supervisors: dr. Carlas Smith, Dylan Kalisvaart

Abstract: Fluorescence microscopy is used for visualizing and studying biological structures at the microscopic level, enabling researchers to achieve nanoscale imaging precision. However, this is limited by the fundamental diffraction limit that affects the resolution in optical microscopy, determining how structures can be distinguished. Fluorescence microscopy techniques that surpass the diffraction limit include structured illumination techniques for applying patterned light, as well as single-molecule localization using stochastic fluorophore activation. Hybrid methods like MINFLUX are able to increase the precision of localization. The Cramér-Rao lower bound is used to establish the theoretical limit for uncertainty in localization precision. The Van Trees Inequality is the Bayesian variation of the Cramér-Rao lower bound that incorporates prior information. In iterative modulation-enhanced single-molecule localization microscopy, the Van Trees Inequality is preferred because it leverages prior knowledge from previous iterations. The current precision laws in iterative modulation-enhanced single-molecule localization microscopy do not perform well under non-ideal conditions typically found in microscopy. So, the optimal light patterns and algorithm parameters for achieving maximum localization precision remain unknown.