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Proceedings Paper

Fluorescence lifetime imaging to differentiate bound from unbound ICG-cRGD both in vitro and in vivo
Author(s): Paulien L. Stegehuis; Martin C. Boonstra; Karien E. de Rooij; François E. Powolny; Riccardo Sinisi; Harald Homulle; Claudio Bruschini; Edoardo Charbon; Cornelis J. H. van de Velde; Boudewijn P. F. Lelieveldt M.D.; Alexander L. Vahrmeijer M.D.; Jouke Dijkstra; Martijn van de Giessen
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Paper Abstract

Excision of the whole tumor is crucial, but remains difficult for many tumor types. Fluorescence lifetime imaging could be helpful intraoperative to differentiate normal from tumor tissue. In this study we investigated the difference in fluorescence lifetime imaging of indocyanine green coupled to cyclic RGD free in solution/serum or bound to integrins e.g. in tumors. The U87-MG glioblastoma cell line, expressing high integrin levels, was cultured to use in vitro and to induce 4 subcutaneous tumors in a-thymic mice (n=4). Lifetimes of bound and unbound probe were measured with an experimental time-domain single-photon avalanche diode array (time resolution <100ps). In vivo measurements were taken 30-60 minutes after intravenous injection, and after 24 hours. The in vitro lifetime of the fluorophores was similar at different concentrations (20, 50 and 100μM) and showed a statistically significant higher lifetime (p<0.001) of bound probe compared to unbound probe. In vivo, lifetimes of the fluorophores in tumors were significantly higher (p<0.001) than at the control site (tail) at 30-60 minutes after probe injection. Lifetimes after 24 hours confirmed tumor-specific binding (also validated by fluorescence intensity images). Based on the difference in lifetime imaging, it can be concluded that it is feasible to separate between bound and unbound probes in vivo.

Paper Details

Date Published: 4 March 2015
PDF: 6 pages
Proc. SPIE 9313, Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XIII, 93130O (4 March 2015); doi: 10.1117/12.2078644
Show Author Affiliations
Paulien L. Stegehuis, Leiden Univ. Medical Ctr. (Netherlands)
Martin C. Boonstra, Leiden Univ. Medical Ctr. (Netherlands)
Karien E. de Rooij, Percuros BV (Netherlands)
Leiden Univ. Medical Ctr. (Netherlands)
François E. Powolny, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
Riccardo Sinisi, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
Harald Homulle, Technische Univ. Delft (Netherlands)
Claudio Bruschini, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
Edoardo Charbon, École Polytechnique Fédérale de Lausanne (Switzerland)
Technische Univ. Delft (Netherlands)
Cornelis J. H. van de Velde, Leiden Univ. Medical Ctr. (Netherlands)
Boudewijn P. F. Lelieveldt M.D., Leiden Univ. Medical Ctr. (Netherlands)
Technische Univ. Delft (Netherlands)
Alexander L. Vahrmeijer M.D., Leiden Univ. Medical Ctr. (Netherlands)
Jouke Dijkstra, Leiden Univ. Medical Ctr. (Netherlands)
Martijn van de Giessen, Leiden Univ. Medical Ctr. (Netherlands)

Published in SPIE Proceedings Vol. 9313:
Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XIII
Anita Mahadevan-Jansen; Tuan Vo-Dinh; Warren S. Grundfest M.D.; Quan Liu, Editor(s)

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