Fei Comapny

Till I.d. Gmbh

Martin Schropp

Rainer Uhl

The invention relates to a method and a device for generating microscopic layer images of 3-dimensional fluorescent objects, the layer images being largely freed from interference signals from other planes. A first region of a sample is subject to a regular illumination pattern formed by a plurality of light islands, with spacing such that the excitation intensity outside the island regions approaches zero. The emission signal emitted by the sample is detected to generate a raw image of the region. The raw image being subject to interpolation of first interference signals from regions outside the illumination islands. Generating a 2-dimensional interference signal map from the interpolated interference signals; and, generating an emission image, freed from interference signals, of the object by subtracting the 2-dimensional interference signal map from the detected emission signal.

Method and device for generating microscopic layer images of 3-dimensional fluorescent objects

A microscope has an objective, a light source illuminating a sample over an illumination beam path, an arrangement producing a flat illumination pattern which is structured in both spatial directions on the sample, a surface detector detecting light coming over one picture beam path, an arrangement shifting the illumination pattern on the sample in one displacement direction, and a control unit taking one picture at a time of the light which was detected by the detector as phase picture in different positions of the pattern along the displacement direction and to computationally reconstruct from these phase pictures an overall picture of the illuminated sample region. The displacement direction is oblique to the main axes of symmetry of the illumination pattern and depending on the illumination pattern is chosen such that the number of phase pictures which is necessary for the picture reconstruction corresponds to the theoretically minimally required value.

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Munich, Germany

Martin Schropp