Patent ID: 8263946

Claim:
A method for high-resolution optical scanning of a sample comprising: a first substance that is switchable into at least a first energy state and a second energy state differing from the first energy state in terms of at least one optical property; a second substance that is switchable into at least a first energy state and a second energy state differing from the first energy state in terms of at least one optical property; wherein the first and second substances differ from one another in at least one property and are at least partly the same in at least one other property, said method comprising the method steps of: illuminating a first area of the sample in an excitation step with illumination light of a wavelength of an excitation spectrum of the first and second substances for local generation of the first energy state of the first and second substances; illuminating a second area of the sample extending peripherally around a focus region of the excited first area in a de-excitation step with illumination light of a suitable de-excitation wavelength for generating the second energy state of the first and second substances; detecting emitted light by means of a detection device, said light being emitted from the sample as a result from a decay of remaining first energy state; scanning of the sample such that at least one of the steps of excitation, de-excitation and detection for the first substance is carried out at a different point in time than for the second substance; and using the illumination light for generating the de-excited state of one of the first or second substances simultaneously for generating the excited state of the other one of said second or first substances.