Enhancement of Fluorescence Mediated by Silver Nanoparticles: Implications for Cell Imaging.

Abstract

In this study, we report the surface enhanced fluorescence (SEF) of a biologically important organic dye, fluorescein (FL), by silver nanoparticles (Ag NPs) in an aqueous medium and its implications for human cell imaging. The as-synthesized Ag NPs were characterized by dynamic light scattering (DLS), zeta potential, transmission electron microscopy (TEM), and UV-vis absorption spectroscopic studies. The interaction and aggregation of FL dye with Ag NPs and a cationic surfactant, namely, cetyltrimethylammonium bromide (CTAB), were explored by UV-vis absorption and steady-state and time-resolved fluorescence spectroscopic methods. The distance-dependent fluorescence enhancement of FL due to Ag NPs in the solution was also theoretically correlated by three-dimensional finite-difference time-domain (3D-FDTD) simulation. The plasmonic coupling between neighboring NPs facilitated the augmentation of the local electric field, thereby producing various "hotspots" that influence the overall fluorescence of the emitter. J-type aggregates of FL in the presence of the CTAB micelles and Ag NP mixed solution were confirmed by electronic spectroscopy. The density functional theoretical (DFT) study revealed the electronic energy levels associated with different forms of FL dye in the aqueous solution. Most interestingly, the Ag NP/FL mixed system used in fluorescence imaging of human lung fibroblast cells (WI 38 cell line) showed a significantly stronger green fluorescence signal compared to that of FL after an incubation period of only 3 h. This study confirms that the Ag NP mediated SEF phenomenon of the FL dye is also manifested in the intracellular medium of human cells giving a brighter and more intense fluorescence image. The cell viability test after exposure to the Ag NP/FL mixed system was confirmed by the MTT assay method. The proposed study may have an implication as an alternate approach for human cell imaging with higher resolution and more contrast.