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Fig. 5. Real-time measurements of simultaneous changes of YFP- and CFP-intensities, FRET-ratio and CBF in single human airway epithelial cells. (A) Dually infected and differentiated cells were basolaterally permeabilized and basolaterally perfused. Intensities of CFP (blue) and YFP (green) emissions were measured during CFP excitation simultaneously with CBF (red). FRET ratio (black) was calculated as quotient of the intensities CFP/YFP. Addition of cAMP to the perfusate is indicated with a bar above the traces. (B) Dually infected and differentiated cells were imaged without permeabilization. Intensities of CFP (blue) and YFP (green) emissions were measured during CFP excitation simultaneously with CBF (red). The FRET ratio (black) was calculated during perfusion of the apical surface with sequential solutions containing 1 µM, 10 µM and 20 µM forskolin.
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