Figure 2

Simultaneous capture of GFP and CFP. Stable cell lines expressing either GFP or CFP were co-cultured (left panels) or a stable cell line expressing CFP and GFP-CAAX (membrane targeted – middle panels) or a cell line expressing CFP transiently transfected with GFP-ZYXIN (right panels) were analyzed. Top panels show the fluorescence captured using a 450/80 filter and non-descanned detection. Upper-mid panels show the fluorescence captured using a 515/30 filter and non-descanned detection. Lower-mid panels show the result of subtracting the 450/80 signal from the 515/30 signal. Bottom panels show a merge of the top panels (CFP signal in blue) and the lower-mid panels (GFP signal in green). GFP-CAAX, which generally targets to the membrane can be seen labeling of intracellular structures in the merge and GFP images (middle panel; arrow). All images were taken with a 40× objective and a final magnification of 500× for the right 2 columns and 1000× for the left. Scale bar = 25 μm for the right 2 columns and 10 μm for the left.