Patent ID: 8080383

Claim:
A method for discriminating between and counting at least two populations of biological elements having specific biological characteristics present in a sample, comprising simultaneously marking each said population of biological elements by three different probes which bind or react to predefined biological characteristics which are specific to the biological elements, the three different probes coupled to three different markers which render the biological characteristics detectable upon binding or reaction, wherein two of said markers are overlapping markers (OMs) each having an emission spectrum, said emission spectra overlapping each other and a third non-overlapping marker (nOM) having an emission spectrum not overlapping the spectra of the other two markers; measuring the total quantity of non-overlapping marker (qnOM) in a detection band chosen in the emission spectrum of said non-overlapping marker; measuring the total quantity of overlapping markers (qOM) in a detection band common to the emission spectra of said overlapping markers; establishing a ratio (R) of the total quantity of non-overlapping marker to the total quantity of overlapping markers [ R =( qnOM )/( qOM )]; and establishing a diagram showing the ratio (R) as a function of the total quantity of biological elements marked by the overlapping marker [R=f(qOM)] wherein the diagram provides discrimination of the populations of biological elements which have predefined biological characteristics.