Patent ID: 9234846
Filing Date: 2016-01-12
CPC Classification: G01N,G02B

Claim Text:
1. A high-resolution microscope for two-dimensional or three-dimensional determination of the position of objects, including individual fluorophores, for spatially high-resolution fluorescence microscopy of a sample, said high-resolution being beyond the optical diffraction limit, which sample is marked with marker molecules, which molecules can be activated or switched by a signal such that they can only be excited to emit fluorescent radiation in an activated or switched state, comprising: 1) means for introducing said signal to the sample such that only a subset of marker molecules provided in the sample are activated, with partial areas in the sample existing in which activated marker molecules are at least at a distance from the closest neighboring activated or switched marker molecules, which distance is greater than or equal to a length resulting from a predetermined optic resolution, said predetermined optic resolution including resolution beyond the diffraction limit, 2) means for exciting the activated molecules to emit fluorescent radiation, 3) means for detecting fluorescent radiation with said predetermined optic resolution, and 4) means for generating an individual image of said fluorescent radiation recorded in step 3), with the geometric location of the marker molecules emitting the fluorescence radiation being determined with a spatial resolution exceeding the predetermined optic resolution, with the foregoing steps being repeated several times and a plurality of individual images resulting therefrom being combined to an overall image, means for multi-plane detection by splitting the detection radiation in a Z direction, and at least one of the following: a cylindrical lens arranged in an imaging beam path as an anamorphotic optic arranged in the display radiation path; at least one beam splitter provided to split the detection radiation path into at least two partial radiation paths and the partial radiation paths comprise a spatial off-set in reference to each other in the detection plane; a short pulse laser provided as the activating or switching source for non-linear sample excitation; and a point scanner or a line scanner provided for non-linear excitation with said short pulse laser.