The method and apparatus for the localization of a single dye molecules in fluorescence microscopy

摘要

The present invention relates to a method for obtaining a sub-resolution spatial information of the sample labeled with at least one fluorescent label. In the method, it is obtained localization image data by using a fluorescent localization microscopy. Localization image data, by using the illumination light having an intensity ranging from about 1 kW / cm ² to about 1 MW / cm ² to illuminate the target region of the sample, when illuminated by at least a portion of the plurality fluorescent molecules of the fluorescent-labeled at least a portion of the emitted fluorescence is detected by the information acquisition sensor, and a detection and illumination repeated a plurality of times, including a series of images taken at different time instances. From localized image data, localization information about the location of at least one fluorescent label a fluorescent molecule at least one spatial position is obtained, in each image, at least one spatial direction, one or more the center of gravity of the distribution of the detected fluorescence emission from a single fluorescent molecules of the fluorescent label is determined.