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Validation of Fluorescence Molecular Tomography/Micro-CT Multimodal Imaging In Vivo in Rats



Rats are important preclinical models for studying breast cancer metastasis and bone pathologies. In these research areas, fluorescence molecular tomography (FMT) is commonly applied for quantitative three-dimensional (3D) imaging in mice. However, uncertainties due to strong depth dependency of FMT signal and spatial resolution require a validation study in rats.


FMT performance in rats was assessed based on co-registered FMT/micro-computed tomography (micro-CT) reconstructed volumes obtained from optical phantoms and from models relevant for tumor imaging, bone remodeling and biodistribution analysis of nanoparticles.


FMT reconstructions within 20-mm-thick optical phantoms were accurate (95 ± 11 % recovery), precise (CV ≤ 8 %) and linear (R 2 > 0.9788) over a range of 78–2,500 nM of the near infrared fluorescent agent VivoTag 750 (VT750). In vivo, implanted defined fluorescent targets yielded a recovery of 105 ± 5 % and successfully co-registered with micro-CT delineated structures. Additionally, using the bone-targeting imaging agent Osteosense 750, regions of neo bone formation identified by FMT could be mapped to the region of epiphyseal growth plates observed in micro-CT images. Finally, as a proof of concept, to monitor nanoparticulate drug pharmacokinetics in rat subjects the accumulation/clearance of VT750–albumin conjugate in/from the liver was followed at 11 different time points over a period of 2 weeks by FMT/micro-CT.


FMT imaging has been validated in optical phantoms as well as in 160 g rats, and sequential FMT/micro-CT imaging can be considered as a useful tool for preclinical research in rats.

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The authors would like to thank Dr. Sasha Belenkov and Dr. Wael Yared from Perkin-Elmer (ViSen) for providing valuable suggestions with respect to FMT operations and software. The assistance of Harald Bartolomae and Alex Rossel in the setup of the imaging equipment is appreciated. The authors also wish to thank Markus Heneka from MicroAnalytics, Germany for assistance with micro-CT related issues. This work was supported by the Excellence Initiative of the German Federal and State Governments Grant EXC 294 and in part by the 5th INTERREG Upper Rhine Program (project A21: Nano@MATRIX).

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Correspondence to V. Prasad Shastri.

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Vonwil, D., Christensen, J., Fischer, S. et al. Validation of Fluorescence Molecular Tomography/Micro-CT Multimodal Imaging In Vivo in Rats. Mol Imaging Biol 16, 350–361 (2014).

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Key words

  • Image co-registration
  • Nanomaterials
  • Nanomedicine
  • Bone remodeling
  • Biodistribution