Method for single molecule localization microscopy

A method for using a light microscope having an objective lens (005) with a resolution limit to form a sequence of images of a biological sample for single molecule localization microscopy, comprising the steps:

• a) exciting emitters (002) in parallel in a field of view of the objective lens with excitation light from an at least one first light source (029);
• b) detecting light emitted from the emitters in parallel as a result of the excitation light, with an at least one first detector (030) to acquire an image;
• c) determining approximate positions (022) of the emitters based on the acquired images;
• d) generating regions corresponding to the approximate positions of the emitters, wherein each region (023) is localized at each emitter in a subset, wherein each region has characteristic dimensions comparable to the resolution limit of the microscope;
• e) adapting a first patterned illumination (024) from at least one light source based on the regions, illuminating a plurality of emitters in parallel in subsequent images;
• and
• and f) repeating steps a)-e) to constitute a ‘dynamic masking’ process (013) in a continuous fashion, updating the patterned illumination based on an appearance and disappearance of the individual emitters in the field of view in the acquired images.