Scanning microscope and method of determining point spread function (PSF) of scanning microscope

摘要

A method for operating a scanning microscope and for determining point spread functions, with which sample images are recorded with the scanning microscope. The method includes scanning a sample with at least one illuminating light beam; recording at least one sample image with a detector device during a scan by the illuminating light beam; and comprising the point spread function, with which a sample image is recorded, from the at least one sample image. A detector device having receiving elements is used, where the distance between the receiving elements is smaller than a diffraction disk that generates a sample point on the detector device. Detector signals, generated by means of the receiving elements, are read out for each of the different positions of the illuminating light beam on the sample, as a result of which the scanning of the sample allows the detector signals, which are read out, to generate a plurality of sample images. In this case the point spread functions with respect to the different detector signals are defined in each instance by means of an illumination point spread function and a detection point spread function. With respect to all of the detector signals, a matching illumination point spread function is assumed that is shifted in accordance with the scanning motion for different detector signals. In addition, with respect to all of the detector signals, a matching detection point spread function is assumed that takes account of a spatial offset between the detector elements. The plurality of sample images are used to compute the illumination point spread function and the detection point spread function, and these functions are used to compute the point spread functions with respect to the different detector signals. In addition, the invention also relates to a corresponding scanning microscope.