Patent ID: 8415161

Claim:
A method for determining, in an instrument for analyzing a multiplicity of fluorescent dyes using a multiplicity of detector channels, a spillover into a secondary detection channel of a fluorescent dye that emits in a primary detection channel; wherein the method comprises: a) determining a calibrated spillover value by measuring, with said instrument, emissions in said primary and secondary detection channels of said fluorescent dye and of a reference material, that emits in said primary and secondary detection channels, and calculating said calibrated spillover value by: (i) multiplying the ratio of the emissions of the fluorescent dye by the ratio of the emissions of the reference material, each measured in the primary and secondary detection channels; or (ii) multiplying the ratio of the emissions measured in the primary detection channel by the ratio of the emissions measured in the secondary detection channel; b) measuring with said instrument, using said instrument set to adjusted photodetector gain settings, emissions of said reference material in said primary detection channel to obtain a first reference value and in said secondary detection channel to obtain a second reference value; and c) determining said spillover into said secondary detection channel of said fluorescent dye by multiplying the calibrated spillover value by the ratio of the first and second reference values to thus obtain a spillover value of the fluorescent dye.