Importance

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Abstract

A sample having at least one probe is illuminated with at least one beam of excitation light that is linearly polarized along a first axis, thereby effecting fluorescence emission in at least one spectral band. 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 at least one spectral band. These measurements are represented as a measurement vector M. Since there are various fluorescence sources besides the at least one probe which emit some limited amount of light in the characteristic band of the at least one probe, spectral cross-talk results between the at least one probe and the other fluorescence sources. 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 one of the other fluorescence sources. A flux vector S is calculated according to S=A.sup.-1M, and the fluorescence polarization FP of the at least one probe is calculated from the S values.

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First Claim