Figure 2

Calibration curves under different PMT gains. X-axis: log10 concentration, Y-axis: log10 fluorescence intensity. A: Cy5 dye; B: Cy3 dye. Representative calibration curves are presented in C (Cy5 and Cy3 channels are scanned under the same PMT gain of 700 V) and D (the Cy5 and Cy3 channels are scanned at 700 V and 400 V, respectively). The range of linear response between dye concentration and fluorescence intensity depends on the PMT gain, so does the slope of the linear response range. There is also a difference between Cy5 and Cy3 in terms of the spread and slope of the linear range. Each data point represents the averaged log intensity of the 12 repeats with the same dye concentration.