High-resolution spectrally selective scanning microscopy of a sample

摘要

In a high-resolution spectrally selective scanning microscopy of a sample, the sample is excited with illumination radiation in order to emit fluorescence radiation such that the illumination radiation is bundled into an illumination spot in or on the sample. The illumination spot is diffraction-limited in at least one spatial direction and has a minimum extension in said spatial direction. Fluorescence radiation emitted from the illumination spot is imaged into a diffraction image lying on an image plane in a diffraction-limited manner and is detected with a spatial resolution which resolves a structure of a diffraction image of the fluorescence radiation emitted from the illumination spot. The illumination spot is moved into different scanning positions. An individual image is generated for each scanning position, in a diffraction-limited manner onto a detector. The local channels determine the spatial resolution with which the structure of the diffraction image of the fluorescence radiation emitted from the illumination spot is resolved, and the fluorescence radiation emitted from the illumination spot is spectrally evaluated.