A fluorescence microscope (10) includes a sample illumination beam path including a source (9) for illumination light, a first wave front modulator (24) for providing the focused illumination light (8) with a central intensity minimum, a beam splitter (26) and a second adjustable wave front modulator (34) arranged in a pupil plane (30) of an objective (20). A first detection beam path section including the second wave front modulator (34) and a telescope (11) and ending at the beam splitter (26) coincides with the sample illumination beam path. A separate second detection beam path section includes a detector (38) for luminescence light from a sample. The telescope (11) images a first pupil (31) formed in the pupil plane (30) in a smaller second pupil (32), and transfers a beam of the illumination light (8) collimated in the second pupil (32) into an expanded beam collimated in the first pupil (31).