Fast high-resolution microscopy method and fast high-resolution microscope

A high-resolution microscope for fast microscopy of a specimen containing a fluorescence emitter, comprising:

• an excitation device that is designed to excite the specimen in the widefield with excitation radiation to emit fluorescent radiation, an imaging device that is designed to image the specimen in the widefield with a spatial resolution, wherein the excitation device is designed such that at least some fluorescence emitters are isolated in the imaging in terms so the spatial resolution,
• a localization analysis device that is designed to determine locations for the isolated fluorescence emitters with an accuracy that exceeds the spatial resolution, and
• a control device that is designed to control the excitation apparatus and the imaging apparatus in accordance with the following method:
• fast high-resolution microscopy method of a specimen containing a fluorescence emitter, in which
• fluorescence emitters are used that have a first and a second state, wherein the first state is a bright state in which the fluorescence emitters emit fluorescent radiation on irradiation of excitation radiation, and the second state is a dark state in which the fluorescence emitters do not emit fluorescent radiation on irradiation of excitation radiation, wherein the fluorescence emitters can be brought from the bright state into the dark state by irradiation of excitation radiation and the dark state has a particular lifetime after which the fluorescence emitters return to the bright state spontaneously,
• the specimen and thus the fluorescence emitters are illuminated in the widefield with excitation radiation of an intensity and the fluorescence emitters are thereby excited to blink, such that they switch back and forth between bright state and dark state at a blinking frequency,
• the blinking specimen is imaged in the widefield with a spatial resolution and is detected with a camera,
• the intensity of the excitation radiation is set such that at least some fluorescence emitters are isolated in the individual images in terms of the spatial resolution, and the intensity of the excitation radiation and a refresh rate of the camera are adapted to each other such that the refresh rate with which the individual images are produced is not lower than the blinking frequency, and
• the individual images are subjected to a localization analysis, wherein locations are determined for isolated fluorescence emitters in the individual images with an accuracy that exceeds the spatial resolution.