Sensitized emission analysis is available when the image has at least three channels (donor, acceptor, FRET; corresponding measured signals are D, A, and F). The measured intensities are first corrected for crosstalk, for which the following factors are considered: F_D/D_D; A_D/F_D; D_A/A_A; D_A/F_A; F_A/A_A. The capital letters refer to channels and subscripts refer to the D and A dyes; for instance, F_D/D_D is the F/D signal ratio measured for a donor-only sample. In addition, unequal detection efficiencies of the donor and FRET channels are considered (G factor).

In all cases the FRET values are calculated for every pixel that is considered “valid” and then the mean value is calculated and presented in the table.