1. New Turf for CFP/YFP FRET Imaging of Membrane Signaling Molecules
Jenafer Evans, David T Yue 
Neuron 
Volume 38, Issue 2, Pages (April 2003)
DOI: /S (03)

2. Figure 1
TIRF Microscopy Enables Selective Detection of FRET from Fluorophore-Tagged, Membrane-Associated Molecules
Laser excitation (454 nm) propagates through the immersion oil and coverslip at the critical angle and strikes the coverslip/specimen interface where there is a change in refractive index. This configuration results in total internal reflection of the incident light, but an evanescent field propagates through a thin segment of the specimen within 100 Å of the coverslip. This allows selective excitation of the CFP-tagged channels in this segment, while CFP-tagged channels in deeper regions of the cell remain dormant. FRET in the membrane segment will result in fluorescence emission from YFP-tagged channel subunits, and such YFP fluorescence is selectively captured in the microscope output. This imaging strategy eliminates confounding FRET signals from channels trapped in intracellular compartments.
Neuron  , DOI: ( /S (03) )