In order to verify that the emission observed using a wide-field microscope is indeed associated with the PCP complexes, we obtain fluorescence spectra and decay
curves for an identically prepared structure. The confocal image, in contrast to the wide-field image, consists of bright spots spread over otherwise quite uniform background. We attribute the spots to the emission of the PCP complexes close to the silica nanoparticles, and the background originates from the PCP complexes placed far away from the nanoparticles. The absence of the ring-like structure on the confocal images is a result of much lower numerical aperture of the collection optics (0.5 vs. 1.4), which results in much lower spatial resolution of the AZD6738 concentration experiment. After collecting such a confocal MCC950 image, we measured spectra and decays for several tens of bright spots and compare the result with the data obtained for the areas free of the nanoparticles. An example of the results is displayed in Figure 4. The comparison of the fluorescence spectra measured for the PCP complexes on and off the nanoparticles (Figure 4a) indicates that the coupling with the nanoparticles leaves no effect upon the spectral shape of the emission. The only impact concerns the total fluorescence intensity and the result that is intact with the observations
made by wide-field microscopy. The average enhancement of the fluorescence emission obtained from this comparison selleckchem is equal to 3. Similarly, the transient behavior of the CYTH4 fluorescence intensity is also identical for the PCP complexes placed on and off the silica nanoparticles (Figure 4b). Unchanged lifetimes indicate that the interaction between the nanoparticles and the photosynthetic complexes induces no changes in the radiative properties of the chlorophyll molecules that are responsible for the fluorescence emission. Figure 4 Emission spectra and fluorescence decay curves of the PCP complexes. (a) Emission spectra of the PCP complexes deposited on (red) and off (black) silica nanoparticles. (b) Fluorescence decay curves of PCP deposited
on (red) and off (black) silica nanoparticles. The excitation wavelength for both experiments was 480 nm. The transients are normalized, and the one measured for the PCP complexes off the silica nanoparticles was shifted vertically (multiplied by 10) for clarity. Conclusions We find that coupling of photosynthetic, chlorophyll-containing complexes with dielectric silica nanoparticles leads to an enhancement of the fluorescence emission. The interaction leaves no measurable effect on the shape of the emission as well as on the transient behavior of the fluorescence. We conclude that the effect of fluorescence enhancement originates from high scattering of electromagnetic field by dielectric nanoparticles that leads to improvement of the collection efficiency.