Suppose we want to add the missing FRET-pairs shown in the lower left corner. These pairs seem to have been missed by the FRET-pair finder due to aberration effects of the optical setup.
First add the donor peak. Select 'Point at donor peaks manually' in the toolbar. This activates a selection tool. Move the tool to the green peak and press left mouse button. Do this for all the D peaks. When finished, press the right mouse button.
Do the same for the acceptors.
Now add the FRET-pairs. Select 'Point at FRET-pairs manually' in the toolbar. With the activated tool, point at the donor and acceptor peak of each FRET-pair, in that order. Finish by right click.
The FRET-pairs have now been added manually.
Remove FRET-pairs interactively in overlay image
To remove D peaks manually, activate the 'Delete donor peaks manually' selection tool in the toolbar. Move the cursor to the desired peaks and press left mouse button. This removes the peak and FRET-pairs containing the selected peak.
Do the same for acceptor peaks.
Use lasso-selection to remove many peaks
If you want to remove all peaks in an area use the lasso tool.
Activate the lasso tool in the toolbar. Move the cursor to the overlay image and drag in the image while pressing down left mouse button.
When finished all peaks within the lasso selection are removed.