Method of spatially measuring a plurality of nano-scale structures in a sample, comprising: - marking the individual structures at different locations with fluorescent markers (8, 9) which are selected from - fluorescent markers (8, 9) that differ in their excitation spectrum and/or their emission spectrum, and - fluorescent markers (8, 9) that have an active state in which they are excitable with excitation light for emission of fluorescence light and an inactive state in which they are, at least with the same excitation light, not excitable for emission of fluorescence light, and that are transferable with switching light between their active and their inactive state, - exciting the fluorescent markers (8, 9) with excitation light for emission of fluorescence light, wherein an intensity distribution of the excitation light or an intensity distribution of further light which has an influence on the emission of fluorescence light by the fluorescent markers (8, 9) comprises a local minimum which is arranged at different positions in the sample, - registering the fluorescence light emitted out of the sample separately for the individual fluorescent markers (8, 9) and for the different positions of the minimum (20, 21, 22, 23), - determining positions of the individual fluorescent markers (8, 9) in the sample from the intensities of the fluorescence light registered for the respective fluorescent marker (8, 9) for the different positions (20, 21, 22, 23) of the minimum, characterized by - coupling the individual structures to individual positioning aids whose positions in the sample are known and/or are determined after the coupling from light reflected by the positioning aids, and - arranging the local minimum at the different positions (20, 21, 22, 23) in a close-up range around the position of the respective positioning aid, whose dimensions are not larger than the diffraction limit at the wavelength of the excitation light and the wavelength of the fluorescence light. - marking the individual structures at different locations with fluorescent markers (8, 9) which are selected from - fluorescent markers (8, 9) that differ in their excitation spectrum and/or their emission spectrum, and - fluorescent markers (8, 9) that have an active state in which they are excitable with excitation light for emission of fluorescence light and an inactive state in which they are, at least with the same excitation light, not excitable for emission of fluorescence light, and that are transferable with switching light between their active and their inactive state, - fluorescent markers (8, 9) that differ in their excitation spectrum and/or their emission spectrum, and - fluorescent markers (8, 9) that have an active state in which they are excitable with excitation light for emission of fluorescence light and an inactive state in which they are, at least with the same excitation light, not excitable for emission of fluorescence light, and that are transferable with switching light between their active and their inactive state, - exciting the fluorescent markers (8, 9) with excitation light for emission of fluorescence light, wherein an intensity distribution of the excitation light or an intensity distribution of further light which has an influence on the emission of fluorescence light by the fluorescent markers (8, 9) comprises a local minimum which is arranged at different positions in the sample, - registering the fluorescence light emitted out of the sample separately for the individual fluorescent markers (8, 9) and for the different positions of the minimum (20, 21, 22, 23), - determining positions of the individual fluorescent markers (8, 9) in the sample from the intensities of the fluorescence light registered for the respective fluorescent marker (8, 9) for the different positions (20, 21, 22, 23) of the minimum, characterized by - coupling the individual structures to individual positioning aids whose positions in the sample are known and/or are determined after the coupling from light reflected by the positioning aids, and - arranging the local minimum at the different positions (20, 21, 22, 23) in a close-up range around the position of the respective positioning aid, whose dimensions are not larger than the diffraction limit at the wavelength of the excitation light and the wavelength of the fluorescence light.