Method for detection of a reversibly photo-convertible fluorescent species

摘要

Method for detection of at least one reversibly photo-convertible fluorescent species (P), comprising the following steps: a) illumination of a sample comprising said or at least one of said reversibly photo-convertible fluorescent species by a periodically modulated illuminating light (FEX); and b) detection of fluorescent radiation (FLU) emitted by the sample thus illuminated; characterised in that the method further comprises the following step: c) extraction of the amplitude (lF0Ut) of the intensity component of said fluorescent radiation exhibiting the same periodicity as said periodically modulated illuminating light and a phase quadrature with respect to the same; and in that the mean intensity of said illuminating light and the modulation frequency of the same are chosen in such a way as to maximise said amplitude of the intensity component of said fluorescent radiation.