LIGHT-MICROSCOPIC METHOD OF LOCALIZATION MICROSCOPY FOR LOCALIZING POINT OBJECTS

摘要

A description is given of a light-microscopic method of localization microscopy for localizing point objects (26) in a sample (14), wherein the sample (14) is imaged onto a detector (16) by means of an imaging optical unit (12); and the point objects (26) contained in the sample (14) are localized within the depth of field range (18) by virtue of the fact that a sample image generated by the imaging of the sample (14) on the detector (16) is taken as a basis for determining lateral x/y positions of the point objects (26) in a direction perpendicular to the optical axis (O), wherein the depth of field range (18) is displaced in the object space relative to the sample (14) along the optical axis (O) at least once by a predetermined axial z-travel distance (△z) and, with axially displaced depth of field range (18), the sample (14) is imaged again onto the detector (16) and a further sample image is generated; the lateral x/y positions of the point objects (26) are determined again on the basis of the further sample image; lateral x/y position deviations between the lateral x/y positions of respectively the same point objects (26) determined on the basis of the different sample images are determined; and correction information is generated depending on the lateral x/y position deviations determined, the lateral x/y positions of the point objects (26) being corrected with the aid of said correction information.