Method, device and non-transitory computer-readable medium for localizing individual emitters in a sample

Abstract

The invention relates to a method for localizing individual emitters in a sample, comprising a pre-localization comprising an illumination of the sample with illumination light, wherein the illumination light induces or modulates light emissions of an individual and stationary emitter in the sample, detecting the light emissions of the emitter and estimating the position of the emitter in the sample from the detected light emissions and a subsequent main localization comprising illuminating the sample with an intensity distribution of the illumination light at illumination positions, the intensity distribution comprising a local minimum, detecting the light emissions of the emitter for the illumination positions, and determining the position of the emitter in the sample from the light emissions detected for the illumination positions, wherein the illumination positions are arranged in a first iteration about the estimated position and wherein the illumination positions in at least one second iteration are arranged around the position of the emitter determined in the preceding iteration, wherein the illumination positions in the first iteration and the at least one second iteration form a respective illumination pattern comprising a maximum extension, wherein the maximum extension of the illumination pattern is kept constant during the main localization. The invention further relates to a light microscope for carrying out the method, a computer program and the use of a fiber bundle for the method.