Sted - fluorescence microscopy with two photons - excitation

摘要

A method for the spatially high-resolution imaging a with a fluorescent dye marked structure in a sample, and pulsed excitation light is focused in the sample, in order to the fluorescent dye located in a focal area for the spontaneous emission of fluorescence light, where the wavelength of the excitation light (5) is selected so that the excitation light (5) by means of a multiphoton process the fluorescent dye or de-excitation light of a different wavelength than the excitation light, is directed onto the sample, in order to the outside of the excited fluorescent dye with respect to the focus region in front of its measuring range is reduced to spontaneous emission, rain, and wherein from the fluorescent dye spontaneously emitted fluorescent light is registered, characterized in that the de-excitation light (10), the a shorter wavelength than the excitation light (5), by means of a plurality of pulses (19) of the excitation light (10) for continuously to the sample (2) is directed.