Method for localization microscopic examination of a sample stained with multiple dyes comprising: - a first acquisition of image data of the sample or a partial area (101,102,103) of the sample, - a determining and selecting a sample region of interest based on the image data, - an exciting of fluorophores in a first subregion within the selected sample region of interest with excitation light of a first excitation wavelength, and a first detecting of the fluorescence photons emitted by the excited fluorophores in a first wavelength region with a first detection unit, wherein a localizing or a tracking of isolated fluorophores is performed in the sample region of interest according to a MINFLUX method or a STED-MINFLUX method, - a second detecting of the fluorescence photons emitted by fluorophores excited with excitation light of the first excitation wavelength in a second wavelength range different from the first wavelength range with a detection unit or the detection unit, characterized in that the first detection unit is configured such that the wavelength range in which it detects fluorescence photons is adjustable, and in that the method further comprises: - a determining of wavelength ranges for an optimized ratiometric separation of the multiple dyes in a localization microscopic image acquisition of the sample in multiple spectral detection channels, wherein the detecting of the fluorescence photons is performed respectively in the wavelength ranges determined for optimized ratiometric separation of the plurality of dyes, wherein, in the ratiometric separation, a fluorophore is assigned to one of the plurality of dyes by determining a ratio of the numbers of fluorescence photons detected in the wavelength ranges. - a first acquisition of image data of the sample or a partial area (101,102,103) of the sample, - a determining and selecting a sample region of interest based on the image data, - an exciting of fluorophores in a first subregion within the selected sample region of interest with excitation light of a first excitation wavelength, and a first detecting of the fluorescence photons emitted by the excited fluorophores in a first wavelength region with a first detection unit, wherein a localizing or a tracking of isolated fluorophores is performed in the sample region of interest according to a MINFLUX method or a STED-MINFLUX method, - a second detecting of the fluorescence photons emitted by fluorophores excited with excitation light of the first excitation wavelength in a second wavelength range different from the first wavelength range with a detection unit or the detection unit, - a determining of wavelength ranges for an optimized ratiometric separation of the multiple dyes in a localization microscopic image acquisition of the sample in multiple spectral detection channels, wherein the detecting of the fluorescence photons is performed respectively in the wavelength ranges determined for optimized ratiometric separation of the plurality of dyes, wherein, in the ratiometric separation, a fluorophore is assigned to one of the plurality of dyes by determining a ratio of the numbers of fluorescence photons detected in the wavelength ranges.