Nanotheranostics 2021; 5(4):448-460. doi:10.7150/ntno.60295

Research Paper

Carbohydrate based biomarkers enable hybrid near infrared fluorescence and 64Cu based radio-guidance for improved surgical precision

Wenbo Wang1,5#, Anders E. Hansen1,5#, Hongmei Sun3#, Frederikke P. Fliedner2, Andreas Kjaer2, Andreas I. Jensen4,5, Thomas L. Andresen1,5, Jonas R. Henriksen1,5✉

1. Technical University of Denmark, Department of Health Technology, Building 423, 2800 Lyngby, Denmark.
2. Department of Clinical Physiology, Nuclear Medicine & PET and Cluster for Molecular Imaging, Department of Biomedical Sciences, Rigshospitalet and University of Copenhagen, Blegdamsvej 9, DK-2100 Copenhagen, Denmark.
3. School of Bioengineering and Food, Key Laboratory of Fermentation Engineering, (Ministry of Education), Key Laboratory of Industrial Microbiology in Hubei, National '111' Center for Cellular Regulation and Molecular Pharmaceutic, Hubei province Cooperative Innovation Center for Industrial Fermentation, Hubei University of Technology, Wuhan 430068, China
4. Technical University of Denmark, The Hevesy Laboratory, Department of Health Technology, 4000 Roskilde, Denmark
5. Center for Nanomedicine and Theranostics, Technical University of Denmark, 2800 Lyngby, Denmark.
# Contributed equally to the work.

This is an open access article distributed under the terms of the Creative Commons Attribution License ( See for full terms and conditions.
Wang W, Hansen AE, Sun H, Fliedner FP, Kjaer A, Jensen AI, Andresen TL, Henriksen JR. Carbohydrate based biomarkers enable hybrid near infrared fluorescence and 64Cu based radio-guidance for improved surgical precision. Nanotheranostics 2021; 5(4):448-460. doi:10.7150/ntno.60295. Available from

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Increasing numbers of lung tumors are identified at early disease stages by diagnostic imaging in screening programs, but difficulties in locating these during surgical intervention has prevented an improved treatment outcome. Surgical biomarkers that are visible on diagnostic images, and that provide the surgeon with real-time image guidance during the intervention are thus highly warranted to bridge diagnostic precision into enhanced therapeutic outcome. In this paper, a liquid soft tissue marker for near infrared fluorescence and radio-guidance is presented. The biocompatible marker is based on the carbohydrate ester, sucrose acetate isobutyrate, ethanol, and a multifunctional naphthalocyanine dye, which enable near infrared fluorescence image-guided resection at short, medium and long tissue depths. Naphthalocyanine dyes have high quantum yields and may further act as chelators of radionuclides. Upon injection of the liquid marker, a gel-like depot is formed in situ at the site of injection, wherein the fluorescent dye and radionuclide is retained. The radiolabeled markers were optimized for minimal fluorescence quenching and high retention of the positron emission tomography radionuclide 64Cu. The performance of the radiolabeled marker was tested in vivo in mice, where it displayed high photostability over a period of 4 weeks, and high retention of 64Cu for 48 hours. The retention and biodistribution of 64Cu was quantified via PET/CT, and the fluorescence emission by an in vivo imaging system. The presented data demonstrate proof-of-concept for naphthalocyanine markers as multimodal imaging agents that can bridge the precision of diagnostic imaging into surgical interventions.

Keywords: Surgical marker, Near infrared fluorescence, PET/CT, Naphthalocyanine, Cu-64