Standardized residuals as a function of the predicted base 10 logarithm median fluorescence intensity (MFI) for the four emulated datasets based on standard curves fitted using the four different approaches to treat background noise.

Points outside of the range specified by the dashed lines are in red and labeled according to the well location in the 96-well plate. (A) Standard curve fitted using “Ignore” background noise method; (B) standard curve fitted including the background noise as an extra point when fitting the curve, using “Include” background method; (C) standard curve fitted subtracting the background noise using the “Subtract” method; (D) standard curve fitted constraining the lower asymptote to the background noise using the “Constraint” method.