Patent Document ID: 9588328
Application ID: 14112519
Patent Status: 1

Claim One:
1. A method of PAL microscopy, wherein placing a sample comprising fluorescence emitters into a wide-field microscope which has an optical resolution and comprises a spatially resolving detector having a radiation-sensitive detector surface, repeating several times a sequence of an isolation step and an imaging step, wherein each isolation step prepares a set of fluorescing emitters in which each fluorescing emitter is spaced apart from all neighboring fluorescing emitters by a minimum distance which is resolvable by the optical resolution of the wide-field microscope, and each imaging step images the fluorescing emitters by the wide-field microscope and provides a single image which contains a radiation distribution of the emitters of the set prepared in the immediate previous isolation step, wherein a sample area is imaged into an image area on the radiation-sensitive detector surface of the spatially-resolving detector, wherein the image area is smaller than the radiation-sensitive detector surface to obtain a single image, and the image area is shifted across the detector surface between successive imaging steps, with the result that in different imaging steps, the same sample area is imaged onto different, adjacent positions of the radiation-sensitive detector surface in order to determine information about changes in the sample area between different single images, and in each single image a position of a center of the radiation distribution is determined for each prepared emitter with a precision higher than the optical resolution of the wide-field microscope, and the positions of the centers of the prepared emitters in the single images are combined to a total image representing the sample with a resolution higher than the optical resolution of the wide-field microscope.