High-Throughput Quantitative Bioluminescence Imaging for Assessing Tumor Burden

  • Angelina Contero
  • Edmond Richer
  • Ana Gondim
  • Ralph P. Mason
Part of the Methods in Molecular Biology™ book series (MIMB, volume 574)


Bioluminescence imaging (BLI) has emerged during the past 5 years as the preeminent method for rapid, cheap, facile screening of tumor growth and spread in mice. Both subcutaneous and orthotopic tumor models are readily observed with high sensitivity and reproducibility. User-friendly commercial instruments exist and, increasingly, luciferase-expressing tumor cells are available in academic institutions or commercially. There is an increasing literature on routine use of BLI for assessing chemotherapeutic efficacy, drug combinations, dosing, and timing. In addition, BLI may be applied to more sophisticated questions of molecular biology by including specific promoter sequences. This chapter will describe routine methods used to support multiple investigators in our small animal imaging resource.

Key words

Luciferase luciferin charged-coupled device cameras (CCD) IGOR Pro bioluminescence 



Supported in part by grants from the DOD Breast Cancer Initiative (IDEA award DAMD17-03-1-0343), the NIH Cancer Imaging Program (P20 CA86354 and U24 CA126608), and the Simmons Cancer Center. We are grateful to Drs. Li Liu, Robert Bollinger, Jerry Shay, and Peter Antich for bringing the vision of BLI to UT Southwestern.


  1. 1.
    Schaefer, C., Mayer, W. K., Krüger, W., and Vaupel, P. (1993) Microregional distributions of glucose, lactate, ATP and tissue pH in experimental tumours upon local hyperthermia and/or hyperglycaemia. J Cancer Res Clin Oncol 119, 599–608.PubMedCrossRefGoogle Scholar
  2. 2.
    Lundin, A. (2000) In Bioluminescence and Chemiluminescence, Pt C 305 346–370.CrossRefGoogle Scholar
  3. 3.
    Contag, C. H., and Ross, B. D. (2002) It's not just about anatomy: in vivo bioluminescence imaging as an eyepiece into biology. JMRI 16, 378–387.PubMedCrossRefGoogle Scholar
  4. 4.
    Thorne, S. H., and Contag, C. H. (2005) Using in vivo bioluminescence imaging to shed light on cancer biology. Proc IEEE 93, 750–762.CrossRefGoogle Scholar
  5. 5.
    Massoud, T. F., and Gambhir, S. S. (2003) Molecular imaging in living subjects: seeing fundamental biological processes in a new light. Genes Dev 17, 545–580.PubMedCrossRefGoogle Scholar
  6. 6.
    Bhaumik, S., and Gambhir, S. S. (2002) Optical imaging of Renilla luciferase reporter gene expression in living mice. Proc Natl Acad Sci USA 99, 377–382.PubMedCrossRefGoogle Scholar
  7. 7.
    Paroo, Z., Bollinger, R. A., Braasch, D. A., Richer, E., Corey, D. R., Antich, P. P., and Mason, R. P. (2004) Validating bioluminescence imaging as a high-throughput, quantitative modality for assessing tumor burden. Mol Imaging 3, 117–124.PubMedCrossRefGoogle Scholar
  8. 8.
    Bollinger, R. A. (2006), Ph.D., UT Southwestern, Dallas.Google Scholar
  9. 9.
    Wang, W., and El-Deiry, W. S. (2003) Bioluminescent molecular imaging of endogenous and exogenous p53-mediated transcription in vitro and in vivo using an HCT116 human colon carcinoma xenograft model. Cancer Biol Ther 2, 196–202.PubMedGoogle Scholar
  10. 10.
    Cecic, I., Chan, D. A., Sutphin, P., Ray, P., Gambhir, S. S., Giaccia, A. J., and Graves, E. E. (2007) Oxygen sensitivity of reporter genes: implications for preclinical imaging of tumor hypoxia. Mol Imaging Biol 6, 219–228.Google Scholar
  11. 11.
    Karam, J. A., Fan, J., Stanfield, J., Richer, E., Benaim, E. A., Frenkel, E., Antich, P., Sagalowsky, A. I., Mason, R. P., and Hsieh, J. -T. (2007) The use of histone deacetylase inhibitor FK228 and DNA hypomethylation agent 5-Azacytidine in human bladder cancer therapy. Int J Cancer 120, 1795–1802.PubMedCrossRefGoogle Scholar
  12. 12.
    Klerk, C. P., Overmeer, R. M., Niers, T. M., Versteeg, H. H., Richel, D. J., Buckle, T., Van Noorden, C. J., and van Tellingen, O. (2007) Validity of bioluminescence measurements for noninvasive in vivo imaging of tumor load in small animals. Biotechniques 43, 7–13PubMedCrossRefGoogle Scholar
  13. 13.
    Sarraf-Yazdi, S., Mi, J., Dewhirst, M. W., and Clary, B. M. (2004) Use of in vivo bioluminescence imaging to predict hepatic tumor burden in mice. J Surg Res 120, 249–255.PubMedCrossRefGoogle Scholar
  14. 14.
    Kanto, V., Munger, J., and Berry, R. (1994) The CCD Camera Cookbook, Willman-Bell, Inc, Richmond, VA.Google Scholar
  15. 15.
    Dikmen, Z. G., Gellert, G., Dogan, P., Mason, R., Antich, P., Richer, E., Wright, W. E., and Shay, J. E. (2005) A new diagnostic system in cancer research: bioluminescent imaging (BLI). Turk J Med Sci 35, 65–70.Google Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Angelina Contero
    • 1
  • Edmond Richer
    • 1
  • Ana Gondim
    • 1
  • Ralph P. Mason
    • 1
  1. 1.Cancer Imaging Program, UT SouthwesternDallasUSA

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