Fluorescently labelled nanomaterials in nanosafety research: Practical advice to avoid artefacts and trace unbound dye

Abstract

Fluorescence labelling has become a fundamental tool in nanotoxicological research. There are, however, certain drawbacks when dealing with the labelling of nanomaterials. Very often the leaching of dye from the nanomaterial or the presence of unbound dyes in solution leads to the incorrect quantification and localisation of nanomaterials in cells. In this review article we will discuss possible situations, which may give rise to incorrect quantification of the fluorescence associated with nanomaterials and their consequences in the evaluation of the fate of the nanomaterial and its intracellular dose. Issues related to the labelling strategies, dye photostability, impact of the dye on the properties of the nanomaterial surface, and the presence of unbound dye will be discussed. We will also show how Fluorescence Correlation Spectroscopy can be used to trace the presence of free label in solution. In addition, we will discuss the interaction of fluorescence molecules with metallic nanoparticles which can lead to an enhancement or quenching of fluorescence depending on the distance between the dye and the nanoparticle surface. Finally, we will compare the fluorescence emission originating from quantum dots and organic molecules.