Abstract: A method deconvolves a multiplexed fluorescence spectral signal into its component single-color fluorescence spectra emitted by contributing fluorophore types. A spectral database contains a parameter set for each of the component spectra. Optical codes are received as the multiplexed signals. A sliding window technique estimates a number and location of local peaks for the component spectra. Allowable variation ranges are assigned for the corresponding parameter sets. Each multiplexed signal is deconvolved into a sum of parametric values to generate a list of its component spectra. Encoded and received optical codes, in the form of multiplexed signals, are decoded and identified by their component spectra and contributing fluorophore types. A system for deconvolving the multiplexed signals includes a spectral database containing the parameter sets, a detection element receiving the optical codes, and a signal processor encoded to analyze and deconvolve the multiplexed signals.