Laser scanning microscope and method for determining a position of a fluorophore

A laser scanning microscope, comprising:

• a light source configured to form an illumination light beam, the illumination light beam having a transverse light intensity profile comprising an intensity minimum located at a center of the illumination light beam,
• a scanning device configured to scan the illumination light beam so that the center of the illumination light beam moves along a closed trajectory in a target area of a specimen,
• a detector configured to detect fluorescence light emitted by a fluorophore within the target area of the specimen, the fluorophore being excited by the illumination light beam, and
• a processor configured to;
• determine an intensity distribution of the fluorescence light as a function of time,
• fit a periodic function to the intensity distribution of the fluorescence light, and
• determine a position of the fluorophore within the target area based on at least one parameter of the fitted periodic function.