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Abstract:
Spectrally resolved detection of fluorescent probes can be used to
identify multiple labeled target molecules in an unknown mixture. We
study how the spectral shape, the experimental noise, and the number of
spectral detection channels affect the success of identification of
weakly fluorescent beads on basis of their emission spectra. The
proposed formalism allows to estimate the performance of the spectral
identification procedure with a given set of spectral codes on the basis
of the reference spectra only. We constructed a simple prism-based setup
for spectral detection and demonstrate that seven distinct but
overlapping spectral codes realized by combining up to three fluorescent
dyes bound to a single bead in a barcode-based manner can be reliably
identified. The procedure allows correct identification even in the
presence of known autofluorescence background stronger than the actual
signal.
