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Fluorescent imaging in the second near infrared window (NIR II, 1-1.4 {\mu}m)holds much promise due to minimal autofluorescence and tissue scattering. Here,using well functionalized biocompatible single-walled carbon nanotubes (SWNTs)as NIR II fluorescent imaging agents, we performed high frame rate videoimaging of mice during intravenous injection of SWNTs and investigated the pathof SWNTs through the mouse anatomy. We observed in real-time SWNT circulationthrough the lungs and kidneys several seconds post-injection, and spleen andliver at slightly later time points. Dynamic contrast enhanced imaging throughprincipal component analysis (PCA) was performed and found to greatly increasethe anatomical resolution of organs as a function of time post-injection.Importantly, PCA was able to discriminate organs such as the pancreas whichcould not be resolved from real-time raw images. Tissue phantom studies wereperformed to compare imaging in the NIR II region to the traditional NIR Ibiological transparency window (700- 900 nm). Examination of the feature sizesof a common NIR I dye (indocyanine green, ICG) showed a more rapid loss offeature contrast and integrity with increasing feature depth as compared toSWNTs in the NIR II region. The effects of increased scattering in the NIR Iversus NIR II region were confirmed by Monte Carlo simulation. In vivofluorescence imaging in the NIR II region combined with PCA analysis mayrepresent a powerful approach to high resolution optical imaging through deeptissues, useful for a wide range of applications from biomedical research todisease diagnostics.

Deep-Tissue Anatomical Imaging of Mice Using Carbon Nanotube Fluorophores in the Second Near Infrared Window