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AssigneeCambridge Research & Instrumentation, Inc.
InventorClifford C. Hoyt
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A sample having a plurality of probe molecules is illuminated with at least one beam of excitation light that is linearly polarized along a first axis, thereby effecting fluorescence emission in a plurality of spectral bands. The intensity of a first component of fluorescence emission that is polarized along the first axis, as well as the intensity of a second component of fluorescence emission that is polarized along an orthogonal second axis, is measured for each of said plurality of spectral bands. These measurements are represented as a measurement vector M. Since each probe emits some limited amount of light in the characteristic band of another probe, this results in cross-talk between probes. The measurement vector is therefore corrected using an instrument response matrix A, which is generated by measuring the flux output of control samples which each have only a single probe species. A flux vector S is calculated according to S=A−1M, and the fluorescence polarization FP is calculated from the S values.
A method for measuring the fluorescence polarization of a plurality of probes in a single sample, which method compensates for spectral cross-talk among said plurality of probes, said method comprising
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