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The 2−ΔΔCT method was as accurate as using standard curves for the calculation of ratios.
Fig. 2 Scheme of the ROIs for calculation of ratios. 1 (yellow): ventricular system; 2 (green): hypothalamic area; 3 (pink): non-target region; and 4 (orange): whole brain.
Calculation of ratios between cancer and normal were fraction-centric (per LC-MS/MS run).
Regional comparisons were based on calculation of ratios for Region 1 R1) to Region 6 R6).
The regional comparisons were based on the calculation of ratios for Region-1 to Region-6.
One advantage of subtraction MRI is that the technique uses no calculation of ratios or formulas.
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Relative quantification was calculated by extrapolation of the standard curve and calculation of ratio levels compared with β-Actin.
In addition, data normalization and calculation of ratio values was performed using the Feature Extraction software 9.1 from Agilent Technologies.
To avoid a denominator being 0 in calculation of ratio values, we added 1 count to every data point.
This procedure yielded the 2 parameters [PpIX 620]η 620 and [PpIX 634]η 634 enabling the calculation of ratio 620/634.
Numeric values of 10-pixels parts along the stripes were averaged before calculation of ratio images to eliminate the camera noise.
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