| abstract |
The invention relates to a method for high-resolution luminescence microscopy of a sample which is labeled with marker molecules which are activatable in such a way that they can be activated only for the purpose of emitting certain luminescent radiation, the method comprising the following steps: a) repeatedly activating and exciting a subset of the marker molecules present in the sample for emitting luminescence radiation, wherein in the sample at least a portion of the luminescent marker molecules have at least a distance greater than or equal to a length to their closest adjacent luminescent marker molecules resulting from a predetermined optical resolution, b) repeatedly imaging the sample with luminescent label molecules along a depth direction and at the predetermined optical resolution and c) generating individual images from the images from step b), wherein in the individual images the geometric locations of the luminescent marking molecules are determined with a spatial resolution which has been increased above the predetermined optical resolution, wherein the resolution along the depth direction d) the activation and / or excitation in steps a) takes place with a radiation which is introduced into at least two areas, wherein The regions each extend along a plane which is substantially perpendicular to the depth direction, - The areas have a predetermined extent in the depth direction and - The areas are arranged so that they are consecutively seen in the depth direction and overlap only partially, and e) in steps b), separate images of the sample are taken upon activation and / or excitation of each of the at least two regions in order to obtain from the separate images a depth information of the luminescent marker molecules. |