The four dye fluorescence detection strategy is a widely used approach
to automated DNA sequence analysis. An important aspect of data proce
ssing in this approach is the multicomponent analysis to deduce the co
ncentrations of four fluorophores from fluorescence emission intensiti
es at four different wavelengths. This requires knowledge of the corre
ct transformation matrix M. The matrix M is a function both of the flu
orophores employed and the fluorescence detection system. M is typical
ly determined either by a calibration process with individual dyes, or
by choosing four well-separated individual peaks corresponding to the
four different dyes. Both are time-consuming and complicated procedur
es for routine use. An automatic scheme for finding M directly from ra
w sequence data is presented here. This facilitates data analysis and
the underlying algorithm may also find utility in other multispectral
applications.