1. To calculate ratios (quotient of two fluorescence intensities) and display ratio images, you need the Physiology (Dynamics) module.
2. You have a suitable image data set open.
3. You are in the MeanROI view on the Ratio tab (view option).

1. Activate the checkbox Enable Ratio Calculation.
2. In the Method dropdown list, select the Single Wavelength (F/F₀) entry.
3. In the Calculation dropdown list select the channel for calculating the ratio.
4. In the Reference image (Ft₀) setup, define the frames of the time series image from which you want the reference value Ft ₀ to be calculated.
5. Click on the Update button.
6. The ratio values are calculated. The ratio image and a diagram for the ratio values are displayed in the MeanROI view. For very large images (pixels and time points) it might be necessary to use the Cache Ratio Image function on the Ratio tab, as this will eliminate flickering when playing through the images at speed.

1. You have successfully calculated a ratio for a single wavelength dye such as Fluo-4.