| Abstract
| - The fast and reversible on/off switching of the fluorescence emission of the GFP-like fluorescentprotein Dronpa has attracted considerable interest for applications in subdiffraction imaging. In this paperwe study the use of a donut-mode beam in combination with two more overlapping laser beams to increasethe imaging resolution through selective switching to the nonfluorescent photoswitched state. We deviseand run a series of numerical simulations to determine suitable photophysical parameters of prospective,thermally stable photoswitchable molecules, in terms of photoswitching quantum yields, fatigue resistance,and possible presence of transient nonfluorescent states. Many of our findings are applicable to othermeasurements that make use of donut beams, and these guidelines can be used in the synthesis andscreening of novel photoswitchable compounds. We experimentally demonstrate the possibility of obtainingincreased resolution by making use of the efficient and thermally stable Dronpa photoswitching, usingequipment that is commonly available.
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