After selecting a correction method and the control images, the next step is to define the areas of interest and then calculate FRET. The resulting data is stored in a data document. The diagram below provides an example of the controls and function of the Analysis panel for FRET and Batch FRET Analysis.
This provides a method for restricting the FRET image display to avoid distracting artifacts or other image details. Use the Enhance, Identify, Modify, Draw/Erase, and Qualify features in the Measurement Tools to define the areas of interest. Anything outside of the defined area will evaluate to zero during FRET calculations.
The next step is to decide whether to measure to a data document or to a spreadsheet. By default, all of the available FRET measurements are selected when FRET Analysis or Batch FRET is selected. So once the objects or areas of interest have been defined, simply click Measure to DataDoc or Measure to Spreadsheet to collect the data.