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Software Manual 10.8.2 Chapter 10 – Intensity Analyses Generating a Ratio Sequence Ratio Open the Ratio dialog window with the Ratio button or via Measure Ratio… Ratio Settings tab Output field: The selection of the options Image, Kinetics and Calibrated determine if a ratio image sequence, a graph or a calcium concentration image sequence are generated. Any assortment of selections is possible. Background Subtraction field. The option None is of limited value because the unavoidable background intensity leads to skewed results when dividing pixel values. If Constant is selected a fixed background value is subtracted from all pixel intensities in all images of the two channels. The constant for the two channels (340 nm and 380 nm) is set in the respective boxes Value 1 and 235