Assessing the Stability of Fluorescently Encoded Nanoparticles in Lysosomes by Using Complementary Methods

Nanoparticles (NPs) are promising tools in biomedical research. In vitro testing is still the first method for initial evaluation; however, NP colloidal behavior and integrity, in particular inside cells (that is, in lysosomes), are largely unknown and difficult to evaluate because of the complexity of the environment. Furthermore, while the majority of NPs are usually labeled with fluorescent dyes for tracking purposes, the effect of the lysosomal environment on the fluorophore properties, as well as the ensuing effects on data interpretation, is often only sparsely addressed. In this work, we have employed several complementary analytical methods to better understand the fate of fluorescently encoded NPs and identify potential pitfalls that may arise from focusing primary analysis on a single attribute, for example, fluorophore detection. Our study shows that in a lysosomal environment NPs can undergo significant changes resulting in dye quenching and distorted fluorescence signals.