Use Direct FRET (Fluorescence Resonance Energy Transfer) as a method of determining whether two types of molecules are in close proximity.
Before you conduct a FRET experiment, consider using the Colocalization tool to determine whether the molecules that you are studying are close to each other. (Colocalization experiments provide more approximate results than FRET but they are much simpler to prepare and analyze.)
Then use the following process to acquire and analyze FRET data:
Acquiring and Preparing FRET Data - To conduct and analyze FRET experiments, follow the softWoRx FRET protocol when you prepare and acquire the data. After you acquire FRET data, you will need to process it. For most applications, this consists of deconvolving the data. Images that are not deconvolved must be adjusted for photosensor measurements made during acquisition. You may need to also correct for chromatic aberration between channels.
Analyzing FRET Data - After processing the data, use the FRET Analysis tool to calculate crosstalk, calculate FRET, and analyze FRET results.