Multiscale Live Imaging Using Förster Resonance Energy Transfer (FRET) for Evaluating the Biological Behavior of Nanoparticles as Drug Carriers

To develop targeted drug delivery systems using nanoparticles for treating various diseases, the evaluation of nanoparticle behavior in biological environments is necessary. In the present study, the biological behavior of polymeric nanoparticles was directly traced in living mice and cells. The dissociation of nanoparticles was detected by Förster resonance energy transfer (FRET) imaging. DiR and DiD were encapsulated in the nanoparticles for near-infrared FRET imaging, and they were traced using in vivo FRET imaging and intravital FRET imaging at the whole-body and tissue scales, respectively. In vivo FRET imaging revealed that the nanoparticles dissociated over time following intravenous administration. Intravital FRET imaging revealed that the nanoparticles dissociated in the liver and blood vessels following intravenous administration. DiI and DiO were encapsulated in nanoparticles for FRET imaging using confocal microscopy, and they were traced using in vitro FRET imaging in HepG2 cells. In vitro FRET imaging revealed that the nanoparticles dissociated and released fluorescent dyes that distributed in the cell membrane. Finally, live imaging was performed using FRET at the whole-body, tissue, and cellular scales. This method is suitable for obtaining information regarding the biological kinetic properties of nanoparticles and their use in targeted drug delivery.