Method, arrangement and microscope for three-dimensional imaging in microscopy using an asymmetric PSF

A microscope comprising:

• an illumination optical unit with an illumination objective for the illumination of a specimen located on a specimen carrier in a specimen region of a specimen plane by means of an illumination radiation over an illumination beam path;
• wherein
• an optical axis of the illumination objective lies in a plane which forms with a normal of the specimen plane with respect to which the specimen carrier is aligned an illumination angle that differs from zero, and illumination takes place in said plane between the detection objective and the detector in the detection beam path
• a beam-shaping means in the illumination beam path for producing a light sheet of the illumination radiation;
• a detection optical unit with a detection objective in a detection beam path extending along an optical axis of which forms with a normal of the specimen plane (4) a detection angle that differs from zero,
• a detection objective for capturing a specimen detection radiation;
• a spatially resolving detector for a two-dimensional capture of image points of the specimen detection radiation;
• and a correction element for the correction of aberrations of the specimen detection radiation between the detection objective and the detector:
• in the detection beam path;
• and a control device for the actuation and controlled setting of the correction element;
• wherein
• the control device is configured to bring about a flawed correction setting on the basis of a best-possible correction setting of the correction element, with which aberrations occurring at the time are reduced as much as possible, in order to produce an asymmetric point spread function of the specimen detection radiation;
• the control device comprises an analysis unit, which is designed and configured for the evaluation of image data captured at the time and the generation of corresponding control commands for actuating actuators of the correction elements to produce said asymmetric point spread function of the specimen detection radiation,
• said control device comprises an analysis unit which is designed and configured for the evaluation of image date captured at the time and the generation of corresponding control commands for actuating actuators of the correction elements.