TY  - GEN
N2  - After circumventing the diffraction limit with super-resolution techniques like STED and PALM/STORM, MINFLUX nanoscopy combines the advantages of these techniques, pushing the optical resolution down to the label size. Thus, more than ever, fluorescence microscopy focuses on single fluorophores, exploiting different fluorescent and non-fluores-cent states. However, the behavior of the photoswitching between these states has not been investigated with respect to the MINFLUX performance. These fluorophore-specific properties heavily influence the image quality, particularly the localization efficiency in terms of success and speed. In this thesis, two dye classes with different excitation-decoupled switching mechanisms (reversible and irreversible) were evaluated for their potential for MINFLUX imaging. To this end, a versatile microscope setup was designed, enabling characterizations on single molecule level in confocal and wide-field illumination mode as well as PALM/STORM microscopy to identify the best dye candidates for MINFLUX imaging. Spontaneously blinking fluorophores (reversible switching) allowed for an up to 30-fold increase of localization speed as compared to previously reported blinking dyes. With photoactivatable dyes (irreversible switching), the localization success of MINFLUX nanoscopy was quantitatively benchmarked for the first time against PALM microscopy. Moreover, intra-marker distances were visualized, provoking a new perspective on labeling strategies for biostructural imaging.
UR  - https://archiv.ub.uni-heidelberg.de/volltextserver/32661/
A1  - Remmel, Michael
ID  - heidok32661
TI  - Characterization of Fluorescent Molecular Switches for MINFLUX Nanoscopy
Y1  - 2023///
AV  - public
CY  - Heidelberg
ER  -