Wide field of view, real time bioimaging apparatus for noninvasive analysis of nanocarrier pharmacokinetics in living model animals.

Abstract

Understanding nanocarrier pharmacokinetics is crucial for the emerging nanopharmacy, which highly demands noninvasive and real-time visualization of the in vivo dynamics of nanocarriers. To this end, we have developed a 2-photon excitation and time-resolved (TPE-TR) bioimaging apparatus for the analysis of the spatial distribution and temporal evolution of nanocarriers in living model animals. The specific polymeric nanocarrier, Eu@pmma-maa doped with Eu-complexes luminescing in long persistence at ∼615 nm upon near-infrared 2-photon excitation, allows the complete rejection of tissue autofluorescence by selective luminescence detection. This together with a unique beam shaping scheme for homogeneous line excitation, a delicate timing strategy for single-shot line scanning, and an equal optical path design for in-plane scan endows the TPE-TR apparatus with the following prominent features: an imaging depth of ∼10 mm, a field of view (FOV) of 32 × 32 mm2 along with a horizontal resolution of ∼60 μm, a sub-10 s frame time, and negligible laser heating effect. In addition, a combination of the in-plane line scan with the 3D scan of a model animal offers the convenience for examining an interested FOV with a millimeter vertical resolution. Application of TPE-TR bioimaging to a living mouse reveals rich information on the dynamics of nanocarriers including the spatial distribution and temporal evolution and the kinetics of domains of interest. The noninvasive TPE-TR bioimaging instrumentation with a wide FOV and a large imaging depth will find applications in the pharmaceutical development of nanocarriers and relevant research fields.