Figure 10

Correction of ratio bias by calculating ratios from the estimated concentrations from the calibration curves. A: Polynomial fitting of c = f(I) for Cy5 at 700 V gain; B: Polynomial fitting of c = f(I) for Cy3 at 700 V gain; C: log standard ratios versus log ratios calculated from concentrations estimated from the calibration curves (Cy5/Cy3 PMT gains at 700 V/700 V); D: log standard ratios versus log ratios calculated from concentrations estimated from the calibration curves (Cy5/Cy3 PMT gains at 400 V/400 V); E: log ratios calculated from concentrations estimated from the calibration curves under Cy5/Cy3 PMT gains at 400 V/700 V and 700 V/400 V, respectively. Data were from the scanner calibration slide. The effectiveness of concentration-based ratio calculation is more obvious by comparing Figures 10C, 10D and 10E to Figures 5B, 5A and 5F, respectively.