A method and a device for cross-talk correction of measured intensities

Abstract

A method for cross-talk correction of intensities measured on mutually separate detection wavelength bands is presented. Each detection wavelength band relates to one of analyte-specific probe-populations contained by a sample to be analyzed. Each probe-population is capable of emitting a first signal component and a second signal component whose spectra have maxima at different wavelengths and at least the first signal component is dependent on presence of analyte detectable with that probe-population. Cross-talk corrected intensities are computed on the basis of a) the intensities measured on the detection wavelength bands, b) a value indicative of intensity occurring on an auxiliary wavelength band outside the detection wavelength bands and at least partially caused by the second signal components, and c) pre-determined cross-talk parameters. For example in con- junction with FRET-based assays, the dependency of a background signal on the percentage of hybridized probes can be taken into account in the cross-talk correction.

Description

technical field [0001] This document discloses methods for crosstalk calibration of intensities measured from a sample over two or more detection bands. Furthermore, the present disclosure relates to devices and computer programs for crosstalk calibration of intensities measured from a sample at two or more detection bands. Additionally, the present disclosure relates to measurement instruments that provide crosstalk calibration. Background technique [0002] A widely used principle in multi-analyte assays is the use of labeled analyte-specific tracer molecules, where detection can be based on changes in the fluorescence-based emission signal that can depend on the excitation wavelength or wavelengths used. The concentration of the analyte in the sample. Ideally, the fluorescence-based emission signals associated with different analytes occur in non-overlapping detection bands, so that the emission signals can be separated from each other using simple optical filtering. H...

AI Technical Summary

Problems solved by technology

The receptor emission signals of different probes hybridized to different target analytes can be separated from each other by using appropriate detection bands, but, as mentioned above, crosstalk complicates the situation

The situation is further complicated by the fact that background emission due to stimulated donors not involved in energy transfer depends on the percentage of hybridized probes to total probes in the considered sample

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