International Journal of Biomedical Imaging

International Journal of Biomedical Imaging / 2006 / Article

Open Access

Volume 2006 |Article ID 037470 |

Abu-Bakr Al-Mehdi, Mita Patel, Abu Haroon, Darla Reed, B. Ohlsson-Wilhelm, K. Muirhead, Brian D. Gray, "Increased Depth of Cellular Imaging in the Intact Lung Using Far-Red and Near-Infrared Fluorescent Probes", International Journal of Biomedical Imaging, vol. 2006, Article ID 037470, 7 pages, 2006.

Increased Depth of Cellular Imaging in the Intact Lung Using Far-Red and Near-Infrared Fluorescent Probes

Academic Editor: Yue Wang
Received19 Jul 2005
Accepted18 Oct 2005
Published08 Mar 2006


Scattering of shorter-wavelength visible light limits the fluorescence imaging depth of thick specimens such as whole organs. In this study, we report the use of four newly synthesized near-infrared and far-red fluorescence probes (excitation/emission, in nm: 644/670; 683/707; 786/814; 824/834) to image tumor cells in the subpleural vasculature of the intact rat lungs. Transpelural imaging of tumor cells labeled with long-wavelength probes and expressing green fluorescent protein (GFP; excitation/emission 488/507 nm) was done in the intact rat lung after perfusate administration or intravenous injection. Our results show that the average optimum imaging depth for the long-wavelength probes is higher (27.8±0.7 μm) than for GFP (20±0.5 μm; p=0.008; n=50), corresponding to a 40% increase in the volume of tissue accessible for high-resolution imaging. The maximum depth of cell visualization was significantly improved with the novel dyes (36.4±1 μm from the pleural surface) compared with GFP (30.1±0.5 μm; p=0.01; n=50). Stable binding of the long-wavelength vital dyes to the plasma membrane also permitted in vivo tracking of injected tumor cells in the pulmonary vasculature. These probes offer a significant improvement in the imaging quality of in situ biological processes in the deeper regions of intact lungs.


  1. A. B. Al-Mehdi, K. Tozawa, A. B. Fisher, L. Shientag, A. Lee, and R. J. Muschel, “Intravascular origin of metastasis from the proliferation of endothelium-attached tumor cells: a new model for metastasis,” Nature Medicine, vol. 6, no. 1, pp. 100–102, 2000. View at: Google Scholar
  2. J. W. Kim, C. W. Wong, J. D. Goldsmith et al., “Rapid apoptosis in the pulmonary vasculature distinguishes non-metastatic from metastatic melanoma cells,” Cancer Letters, vol. 213, no. 2, pp. 203–212, 2004. View at: Google Scholar
  3. H. Wang, W. Fu, J. H. Im et al., “Tumor cell a3ß1 integrin and vascular laminin-5 mediate pulmonary arrest and metastasis,” The Journal of Cell Biology, vol. 164, no. 6, pp. 935–941, 2004. View at: Google Scholar
  4. B. W. Rice, M. D. Cable, and M. B. Nelson, “In vivo imaging of light-emitting probes,” Journal of Biomedical Optics, vol. 6, no. 4, pp. 432–440, 2001. View at: Google Scholar
  5. B. Mohlenhoff, M. Romeo, M. Diem, and B. R. Wood, “Mie-type scattering and non-Beer-Lambert absorption behavior of human cells in infrared microspectroscopy,” Biophysical Journal, vol. 88, no. 5, pp. 3635–3640, 2005. View at: Google Scholar
  6. J. D. Wilson and T. H. Foster, “Mie theory interpretations of light scattering from intact cells,” Optics Letters, vol. 30, no. 18, pp. 2442–2444, 2005. View at: Google Scholar
  7. A. Wunder, C. H. Tung, U. Muller-Ladner, R. Weissleder, and U. Mahmood, “In vivo imaging of protease activity in arthritis: a novel approach for monitoring treatment response,” Arthritis & Rheumatism, vol. 50, no. 8, pp. 2459–2465, 2004. View at: Google Scholar
  8. R. Weissleder, C. H. Tung, U. Mahmood, and A. Bogdanov Jr., “In vivo imaging of tumors with protease-activated near-infrared fluorescent probes,” Nature Biotechnology, vol. 17, no. 4, pp. 375–378, 1999. View at: Google Scholar
  9. V. Ntziachristos, C. Bremer, and R. Weissleder, “Fluorescence imaging with near-infrared light: new technological advances that enable in vivo molecular imaging,” European Radiology, vol. 13, no. 1, pp. 195–208, 2003. View at: Google Scholar
  10. N. Y. Morgan, S. English, W. Chen et al., “Real time in vivo non-invasive optical imaging using near-infrared fluorescent quantum dots(1),” Academic Radiology, vol. 12, no. 3, pp. 313–323, 2005. View at: Google Scholar
  11. N. Bercovici, A. L. Givan, M. G. Waugh et al., “Multiparameter precursor analysis of T-cell responses to antigen,” Journal of Immunological Methods, vol. 276, no. 1-2, pp. 5–17, 2003. View at: Google Scholar
  12. R. Y. Poon, B. M. Ohlsson-Wilhelm, C. B. Bagwell, and K. A. Muirhead, “Use of PKH membrane intercalating dyes to monitor cell trafficking and function,” in In Living Color: Flow Cytometry and Cell Sorting Protocols, R. A. Diamond and S. DeMagio, Eds., pp. 302–352, Springer, New York, NY, USA, 2000. View at: Google Scholar
  13. A. B. Al-Mehdi, G. Zhao, C. Dodia et al., “Endothelial NADPH oxidase as the source of oxidants in lungs exposed to ischemia or high K+,” Circulation Research, vol. 83, no. 7, pp. 730–737, 1998. View at: Google Scholar
  14. K. Konig, K. Schenke-Layland, I. Riemann, and U. A. Stock, “Multiphoton autofluorescence imaging of intratissue elastic fibers,” Biomaterials, vol. 26, no. 5, pp. 495–500, 2005. View at: Google Scholar
  15. Y. Chen, X. Intes, and B. Chance, “Development of high-sensitivity near-infrared fluorescence imaging device for early cancer detection,” Biomedical Instrumentation & Technology, vol. 39, no. 1, pp. 75–85, 2005. View at: Google Scholar
  16. D. Citrin, T. Scott, M. Sproull, C. Menard, P. J. Tofilon, and K. Camphausen, “In vivo tumor imaging using a near-infrared-labeled endostatin molecule,” International Journal of Radiation Oncology, Biology, Physics, vol. 58, no. 2, pp. 536–541, 2004. View at: Google Scholar
  17. X. Chen, P. S. Conti, and R. A. Moats, “In vivo near-infrared fluorescence imaging of integrin αvβ3 in brain tumor xenografts,” Cancer Research, vol. 64, no. 21, pp. 8009–8014, 2004. View at: Google Scholar
  18. A. Hansch, O. Frey, D. Sauner et al., “In vivo imaging of experimental arthritis with near-infrared fluorescence,” Arthritis & Rheumatism, vol. 50, no. 3, pp. 961–967, 2004. View at: Google Scholar
  19. E. M. Gill, G. M. Palmer, and N. Ramanujam, “Steady-state fluorescence imaging of neoplasia,” Methods in Enzymology, vol. 361, pp. 452–481, 2003. View at: Google Scholar
  20. A. Petrovsky, E. Schellenberger, L. Josephson, R. Weissleder, and A. Bogdanov Jr., “Near-infrared fluorescent imaging of tumor apoptosis,” Cancer Research, vol. 63, no. 8, pp. 1936–1942, 2003. View at: Google Scholar
  21. J. V. Frangioni, “In vivo near-infrared fluorescence imaging,” Current Opinion in Chemical Biology, vol. 7, no. 5, pp. 626–634, 2003. View at: Google Scholar
  22. A. Eidsath, V. Chernomordik, A. Gandjbakhche, P. Smith, and A. Russo, “Three-dimensional localization of fluorescent masses deeply embedded in tissue,” Physics in Medicine and Biology, vol. 47, no. 22, pp. 4079–4092, 2002. View at: Google Scholar

Copyright © 2006 Abu-Bakr Al-Mehdi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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