Abstract: | Introduction: The application of intravenously administered nanoparticle (NP) therapies is increasingly being explored for a variety of diseases. The key to their success lies in a thorough understanding of nanoparticle pharmacological behavior, specificity and elimination kinetics. Fluorescent imaging techniques provide exciting opportunities to gain insight into such NP behavior in complex biological systems, at macroscopic as well as microscopic levels.Areas covered: In this review, we will summarize NP labeling methods in relation to their established and emerging fluorescent imaging modalities for in vitro, in vivo, and ex vivo studies of NP behavior. We will highlight novel applications and discuss recent developments in techniques such as fluorescence molecular tomography (FMT), Förster resonance energy transfer (FRET), and Raman imaging. Finally, we will provide a perspective on the challenges and future directions of NP-based fluorescent labeling and imaging in nanotherapeutics.Expert opinion: Commonly used in preclinical applications, fluorescent imaging of NPs can be achieved with minimal invasiveness and low toxicity in a multiplex fashion. Increasingly applied in the study of NP biodistribution, dissociation, and elimination behavior, fluorescent imaging allows fluid longitudinal tracking and visualization of NP interactions at the level of the whole animal, target organs/tissues, and individual cells. |