FRET - Image analysis - FRETR (Page 4)
The total intensity data for each object imaged is then brought into a spreadsheet. In the image shown columns A - E come directly from the data text file. For the other columns: G=A-B; H=C-D; I=E-F; J=G*YFP Spillover factor (here .232), K=I*CFP Spillover factor (here .446); L=J+K; M=H/L. Finally the average of the FRET R column (M) yields the FRETR. We have a Perl script that automates the whole process for our DeltaVision files, but it can be done manually by cutting and pasting using a text editor and Excel. The interpretation of
FRETR is presented in the next, and final web page of this tutorial.
