METHOD FOR 3D SUPER-RESOLUTION LOCALIZED MICROSCOPY

摘要

PROBLEM TO BE SOLVED: To present a depth-resolution microscopy with which depth information not limited by whether or not fluorescent molecules are located within a predetermined depth range is determined within a wider depth range, and unnecessary irradiation of a fluorescent marker can be avoided as much as possible.SOLUTION: In a method for a 3D super-resolution localized microscopy, an excitation step and an imaging step are repeated multiple times to create a plurality of single images; from separated images of a light-emitting fluorescent marker, positional information on the respective corresponding fluorescent markers having the accuracy beyond optical resolution is determined in the plurality of created single images; the entire image with super resolution is created from thus determined positional information; a sample is irradiated with an excitation radiation as a first optical sheet 4, where the first optical sheet has intensity distribution asymmetric with respect to a focal plane along the imaging direction; the separated images of the light-emitting fluorescent marker are analyzed in terms of the contour 13 in the single images; and information on the distance of the corresponding fluorescent marker from the focal plane is derived from the contour.