Spectral-resolution, high-resolution 3D localization microscopy

摘要

A localization microscope is described which has an imaging device for imaging a sample (2) emitting sample light from a focal plane (3) into an image plane (9), the imaging device having an optical manipulation device (18) which is used to influence a point image washing function as a function of depth of the imaging and influences the point image washing function of the imaging in such a way that a point emitter in the image plane is imaged in a point emitter image (30, 32; 42) that is recorded rotationally asymmetrically, - a shape and / or orientation of the rotationally asymmetrical distortion of the point emitter image ( 30, 32; 42) depend on the position of the point emitter with respect to the focal plane (3) and a wavelength of the sample light and - the optical manipulation device (18) having a first and a second anisotropy element (22, 24), each of which has the point image washing function of the imaging anisotropic to generate the rotational asymmetry of the point emitter image and each have an anisotropy axis (23, 25), characterized in that the two anisotropy elements (22, 24) are arranged one behind the other in the imaging direction and their anisotropy axes (23, 25) at an angle lie to each other and both anisotropy elements (22, 249 each have a neutral wavelength (λ, λ), at which they do not anisotropically influence the point-image washing function of the imaging, - the neutral wavelengths (λ, λ) being different.