Patent ID: 6566143
Filing Date: 2003-05-20
Classification: G01N

Abstract:
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 comprisingilluminating a sample having a plurality of probes with at least one linearly polarized beam of excitation light, thereby effecting fluorescence emission in a plurality of spectral bands, measuring the intensity of a first component of fluorescence emission that is linearly polarized along a first axis, in each of said plurality of spectral bands, measuring the intensity of a second component of fluorescence emission that is linearly polarized along a second axis that is orthogonal to the first axis, in each of said plurality of spectral bands, illuminating each of a plurality of samples which each have only a respective one of said probes with at least one linearly polarized beams of excitation light, thereby effecting fluorescence emission in at least one of said plurality of spectral bands from each of said plurality of samples, measuring the intensities of said first and second components of fluorescence emission in each of said plurality of spectral bands for each of said samples which each have only a respective one of said probes, and compensating for spectral cross-talk amongst said plurality of probes by determining the relative contributions from each of said probes to the measured intensities of said first and second components of fluorescence emission at each spectral band, and calculating the fluorescence polarization of each of said plurality of probes based on said relative contributions.