Patent Document ID: 20140340483
Application ID: 14276167
Patent Status: 0

Claim One:
1. A method for three-dimensional high-resolution localization microscopy comprising: illuminating a sample by excitation radiation in an excitation step so as to excite fluorescence markers in the sample to luminesce; imaging the sample in an image frame in an imaging step by means of imaging optics along an imaging direction, wherein the image frame contains images of the luminescing fluorescence markers, and the imaging optics have a plane of focus and an optical resolution; repeating the excitation step and imaging step multiple times to generate a plurality of image frames; isolating images of the luminescing fluorescence markers in each image frame for at least some of the luminescing fluorescence markers; determining a location of the corresponding fluorescence marker in each instance in the generated plurality of image frames from the isolated images of the luminescing fluorescence markers, which location has an accuracy exceeding the optical resolution; and generating a highly resolved total from the determined locations; wherein the excitation radiation is passed into the sample as a first light sheet having along the imaging direction an intensity distribution that is asymmetric to the plane of focus; and wherein the isolated images of the luminescing fluorescence markers are each analyzed with respect to a contour shape thereof in the image frames, and information about a distance of the corresponding fluorescence marker from the plane of focus is derived from the contour shape.