A method for the three-dimensional imaging of a sample in which image information from different depth planes of the sample is stored in a spatially resolved manner, and the three-dimensional image of the sample is subsequently reconstructed from this stored image information is provided. A reference structure is applied to the illumination light, at least one fluorescing reference object is positioned next to or in the sample, images of the reference structure of the illumination light, of the reference object are recorded from at least one detection direction and evaluated. The light sheet is brought into an optimal position based on the results and image information of the reference object and of the sample from a plurality of detection directions is stored. Transformation operators are obtained on the basis of the stored image information and the reconstruction of the three-dimensional image of the is based on these transformation operators.