Figure 3

Pt -GFP emission signals are markedly higher than unspecific background signals. (A) Overlay of ex 490 nm (pseudo-green) and ex 387 nm (pseudo-blue) fluorescence images shows adaxial leaf apoplast that was partially loaded with the Pt-GFP. Under illumination, dye-loaded areas appear yellowish because pseudo-green and pseudo-blue are mixed within the overlay. Areas that were not loaded with the pH reporter protein appear black when illuminated and serve to compare the amount of unspecific signals (background) to signals that are specifically emitted from the proteins. For this comparison, the intensity of grey values was chosen as a measure for signal intensity. A profile of the intensity values was taken from the white line in (A) and is presented in (B). Profiles are displayed for both fluorescence excitation channels (F₄₉₀ and F₃₈₇). Only negligible signals were emitted from the area without dye and signals were much higher in the dye-loaded areas. Twelve separate images captured from different plants proved the low background intensity.