Microscope and method for high resolution 3D fluorescence microscopy

摘要

In a sample, fluorescence emitters are repeatedly excited to emit fluorescence, and still images are produced of the sample by means of a microscope. At least a subset of the fluorescence emitters is isolated in each still image. The positions of the fluorescence emitters are localized in the still images with a location accuracy exceeding the optical resolution. A high-resolution composite image is generated therefrom. An adaptive mirror is arranged in the imaging beam path, and is adjusted in such a manner that it produces an astigmatism when at least one of the still images is produced. As a result, still images with astigmatism are captured. Depth position information for the fluorescence emitters is derived from the rotational asymmetry. The adaptive mirror is additionally adjusted in such a manner that it does not produce any astigmatism when some of the still images are produced.