Use of Panitumumab-IRDye800 to Image Microscopic Head and Neck Cancer in an Orthotopic Surgical Model



Fluorescence imaging hardware (SPY) has recently been developed for intraoperative assessment of blood flow via detection of probes emitting in the near-infrared (NIR) spectrum. This study sought to determine if this imaging system was capable of detecting micrometastatic head and neck squamous cell carcinoma (HNSCC) in preclinical models.


A NIR fluorescent probe (IRDye800CW) was covalently linked to a monoclonal antibody targeting epidermal growth factor receptor (EGFR; panitumumab) or nonspecific IgG. HNSCC flank (SCC-1) and orthotopic (FADU and OSC19) xenografts were imaged 48–96 h after systemic injection of labeled panitumumab or IgG. The primary tumor and regional lymph nodes were dissected using fluorescence guidance with the SPY system and grossly assessed with a charge-coupled NIR system (Pearl). Histologic slides were also imaged with a NIR charged-coupled device (Odyssey) and fluorescence intensity was correlated with pathologic confirmation of disease.


Orthotopic tongue tumors were clearly delineated from normal tissue with tumor-to-background ratios of 2.9 (Pearl) and 2.3 (SPY). Disease detection was significantly improved with panitumumab-IRDye compared to IgG-IRDye800 (P < 0.05). Tissue biopsy samples (average size 3.7 mm) positive for fluorescence were confirmed for pathologic disease by histology and immunohistochemistry (n = 25 of 25). Biopsy samples of nonfluorescent tissue were proven to be negative for malignancy (n = 28 of 28). The SPY was able to detect regional lymph node metastasis (<1.0 mm) and microscopic areas of disease. Standard histological assessment in both frozen and paraffin-embedded histologic specimens was augmented using the Odyssey.


Panitumumab-IRDye800 may have clinical utility in detection and removal of microscopic HNSCC using existing intraoperative optical imaging hardware and may augment analysis of frozen and permanent pathology.

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. 1.

    Spaulding DC, Spaulding BO. Epidermal growth factor receptor expression and measurement in solid tumors. Semin Oncol. 2002;29:45–54.

  2. 2.

    Terwisscha van Scheltinga AG, van Dam GM, Nagengast WB, et al. (2011) Intraoperative near-infrared fluorescence tumor imaging with vascular endothelial growth factor and human epidermal growth factor receptor 2 targeting antibodies. J Nucl Med. 52:1778–85.

  3. 3.

    Themelis G, Yoo JS, Soh KS, et al. Real-time intraoperative fluorescence imaging system using light-absorption correction. J Biomed Opt. 2009;14:064012.

  4. 4.

    Keereweer S, Kerrebijn JD, van Driel PB, et al. Optical image-guided surgery—where do we stand? Mol Imaging Biol. 2011;13:199–207.

  5. 5.

    Keereweer S, Kerrebijn JD, Mol IM, et al. Optical imaging of oral squamous cell carcinoma and cervical lymph node metastasis. Head Neck. 2011. doi:10.1002/hed.21861.

  6. 6.

    Biosciences L-C. 2007IRDye(R) 800CW protein labeling kit—high MW. Lincoln, NE: Li-Cor Biosciences; 2007. p. 1–9.

  7. 7.

    Reuthebuch O, Haussler A, Genoni M, et al. Novadaq SPY: intraoperative quality assessment in off-pump coronary artery bypass grafting. Chest. 2004;125:418–24.

  8. 8.

    Gleysteen JP, Newman JR, Chhieng D, et al. Fluorescent labeled anti-EGFR antibody for identification of regional and distant metastasis in a preclinical xenograft model. Head Neck. 2008;30:782–9.

  9. 9.

    van Dam GM, Themelis G, Crane LM, et al. Intraoperative tumor-specific fluorescence imaging in ovarian cancer by folate receptor-alpha targeting: first in-human results. Nat Med. 2011;17:1315–9.

  10. 10.

    Pleijhuis RG, Graafland M, de Vries J, et al. Obtaining adequate surgical margins in breast-conserving therapy for patients with early-stage breast cancer: current modalities and future directions. Ann Surg Oncol. 2009;16:2717–30.

  11. 11.

    Rosenthal EL, Kulbersh BD, Duncan RD, et al. In vivo detection of head and neck cancer orthotopic xenografts by immunofluorescence. Laryngoscope. 2006;116:1636–41.

  12. 12.

    Gleysteen JP, Duncan RD, Magnuson JS, et al. Fluorescently labeled cetuximab to evaluate head and neck cancer response to treatment. Cancer Biol Ther. 2007;6:1181–5.

  13. 13.

    Kulbersh BD, Duncan RD, Magnuson JS, et al. Sensitivity and specificity of fluorescent immunoguided neoplasm detection in head and neck cancer xenografts. Arch Otolaryngol Head Neck Surg. 2007;133:511–5.

  14. 14.

    Marshall MV, Draney D, Sevick-Muraca EM, Olive DM. Single-dose intravenous toxicity study of IRDye 800CW in Sprague-Dawley rats. Mol Imaging Biol. 2010;12:583–94.

  15. 15.

    Adams KE, Ke S, Kwon S, et al. Comparison of visible and near-infrared wavelength-excitable fluorescent dyes for molecular imaging of cancer. J Biomed Opt. 2007;12:024017.

  16. 16.

    Pomerantz RG, Grandis JR. The epidermal growth factor receptor signaling network in head and neck carcinogenesis and implications for targeted therapy. Semin Oncol. 2004;31:734–43.

  17. 17.

    Withrow KP, Gleysteen JP, Safavy A, et al. Assessment of indocyanine green–labeled cetuximab to detect xenografted head and neck cancer cell lines. Otolaryngol Head Neck Surg. 2007;137:729–34.

  18. 18.

    Remsen KA, Lucente FE, Biller HF. Reliability of frozen section diagnosis in head and neck neoplasms. Laryngoscope. 1984;94:519–24.

  19. 19.

    Gandour-Edwards RF, Donald PJ, Lie JT. Clinical utility of intraoperative frozen section diagnosis in head and neck surgery: a quality assurance perspective. Head Neck. 1993;15:373–6.

  20. 20.

    Keereweer S, Sterenborg HJ, Kerrebijn JD, et al. Image-guided surgery in head and neck cancer: current practice and future directions of optical imaging. Head Neck. 2012;34:120–6.

  21. 21.

    Zijlstra A, Mellor R, Panzarella G, et al. A quantitative analysis of rate-limiting steps in the metastatic cascade using human-specific real-time polymerase chain reaction. Cancer Res. 2002;62:7083–92.

Download references


Supported in part by a grant from NIDCR (R21DE019232) and equipment donated by Novadaq. The authors thank Yolanda Hartman for running the Western blot assays.

Author information

Correspondence to Eben L. Rosenthal MD.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (M4V 30,045 kb)

Supplementary material 1 (M4V 30,045 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Heath, C.H., Deep, N.L., Sweeny, L. et al. Use of Panitumumab-IRDye800 to Image Microscopic Head and Neck Cancer in an Orthotopic Surgical Model. Ann Surg Oncol 19, 3879–3887 (2012) doi:10.1245/s10434-012-2435-y

Download citation


  • Epidermal Growth Factor Receptor
  • Human Epidermal Growth Factor Receptor
  • Cetuximab
  • Indocyanine Green
  • Neck Squamous Cell Carcinoma