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This study aims to develop a novel cross-sectional imaging of fluorescence in over-1000 nm near-infrared (OTN-NIR), which allows in vivo deep imaging, using computed tomography (CT) system. Cylindrical specimens of composite of OTN-NIR fluorophore, NaGdF4 co-doped with Yb3+ and Ho3+ (ex: 980 nm, em: 1150 nm), were embedded in cubic agar (10.5-12 mm) or in the peritoneal cavity of mice and placed on a rotatable stage. When the fluorescence from inside of the samples was serially captured from multiple angles, the images were disrupted by the reflection and refraction of emitted light on the sample-air interface. Immersing the sample into water filled in a rectangular bath suppressed the disruption at the interface and successfully reconstructed the position and concentration of OTN-NIR fluorophores on the cross-sectional images using a CT technique. This is promising as a novel three-dimensional imaging technique for OTN-NIR fluorescent image projections of small animals captured from multiple angles.

The refraction of fluorescence from the inside of a sample at the surface results in fluctuations in fluorescence computed tomography (CT). We evaluated the influence of the difference in refractive index (RI) between the sample body and the surroundings on fluorescence CT results. The brightest fluorescent point is away from the correct point on the tomograms owing to the refraction. The speculated position is determined as the exact point if the RI ratio ranges between 0.97 and 1.03 by immersing the body in an RI matching liquid. The results can help in experimental settings of fluorescence CT for acquiring three-dimensional positional information.