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Abstract

The subject matter of the invention is a method for measuring the luminescence of a long-life fluorescent compound present in a measuring medium, said medium containing a biological sample, characterized in that it comprises the following steps:

a) introduction of a long-life fluorescent compound into the measuring medium,b) introduction, into the measuring medium, of a fluorescent marking agent, the absorption spectrum of which allows the excitation thereof at the same wavelength as that used to excite the long-life fluorescent compound, wherein said emission spectrum allows the measurement of the luminescence thereof at the same wavelength as that used to measure the luminescence of the long-life fluorescent compound,c) excitation of the measuring medium with a wavelength corresponding to an absorption peak of the long-life fluorescent compound,d) measurement of the luminescence emitted by the measuring medium immediately after excitation of said medium, mainly corresponding to the luminescence of the marking agent, and for a period of 5 ns to 45 μs, at a wavelength correspondant to an emission peak of the long-life fluorescent compound,e) time-resolved measurement of the luminescence emitted by the measuring medium at the same wavelength as that used in step d), after a delay of 20 to 200 μs following the excitation of the measuring medium and for a period of 200 to 1000 μs, this luminescence mainly corresponding to that of the long-life fluorescent compound,f) calculation of a normalized luminescence signal corresponding to the ratio: (signal obtained in step e)/(signal obtained in step d).

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