Method and device for high-resolution fluorescence microscopy

Abstract

A method for high-resolution fluorescence microscopy, in which a number N of partial images of a specimen marked with fluorophores and excited to emit fluorescence are recorded, wherein the specimen is successively illuminated by N different effective illuminating patterns, a composite image is calculated from the partial images, the composite image having a higher structural resolution than the partial images, and the composite image is subsequently output, wherein each effective illuminating pattern is generated from the superimposition of at least two basic illuminating patterns, with the basic illuminating patterns differing from each other and for each partial image, and wherein the fluorophores contained in the sample are excited to emit fluorescence only where the at least two basic illuminating patterns superimpose to illuminate the sample, with the illumination superimposition of the at least two basic illuminating patterns releasing non-linear excitation and/or emission effects and/or switching effects in the fluorophores.

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