Fig. 8. FRET assay for AtMinD1 protein-protein interaction in chloroplasts. AtMinD1-CFP and/or AtMinD1-YFP were expressed in tobacco leaves by particle bombardment. Fluorescence of CFP and YFP in leaf epidermal chloroplasts was detected by epifluorescence microscopy. In the FRET channel, emission of YFP was detected upon CFP excitation. (A) Single and dual expression of fluorescent protein-tagged AtMinD1 proteins. (B) Moderate photobleaching of the acceptor YFP leading to the increased emission of CFP. All fluorescent images were taken at the same exposure time (200 milliseconds) using a 60x objective lens (Nikon). Emission signals of CFP (light blue) and YFP (yellowish green) are pseudo-colored. Bars, 5 µm (A) and 1 µm (B).