Bioluminescence Imaging


Bioluminescence imaging (BLI) has emerged over the last decades as a non-invasive assessment of a molecular target. A luciferase reporter gene is expressed in cells of interest and the enzymatic turnover of luciferase after administration of its substrate allows the readout of the reporter gene activity. The signal intensity reflects the strength or changes of a molecular target in a semi-quantitative way.
Depending on the research question at hand, different strategies can be used.
The reporter gene can be expressed constitutively, to monitor the survival and cell growth of transplanted cells in vivo, e.g. stem cells or cancer cells. Integration of luciferase reporters into pathogens or vectors, such as bacteria, viruses and parasites, enables the assessment and follow-up of in vivo infection and its therapeutic response. A second strategy is to use a non-constitutive promoter to monitor gene expression. Luciferase expression is driven by promoter sequences from the genes of interest or placed downstream of genetic elements responsive to a chosen transcription factor. Using this approach, BLI readout will report the up- or down-regulation of this pathway.

ICMI lab has performed BLI in the following research settings:

  • Optimization of BLI and identification of influencing factors
BLI Figuur IV_IP_KeyaertsM

A number of influencing factors have been identified and have resulted in multiple papers, including a review paper.Comparison of the signal intensity using intraperitoneal (IP) and intravenous injection of the substrate D-luciferin in the same mouse. The tumor on the left side of the mouse has a higher light emission after IV injection. The tumor on the right side is only clearly discernable from background after IV injection, indicating the higher sensitivity using IV injection.

Keyaerts M, Caveliers V, Lahoutte T. Bioluminescence imaging: looking beyond the light. Trends Mol Med 2012.

Keyaerts M, Remory I, Caveliers V, Breckpot K, Bos TJ, Poelaert J, Bossuyt A, Lahoutte T. Inhibition of firefly luciferase by general anesthetics: effect on in vitro and in vivo bioluminescence imaging. PLoS One 2012; 7: e30061.

Keyaerts M, Heneweer C, Gainkam LO, Caveliers V, Beattie BJ, Martens GA, Vanhove C, Bossuyt A, Blasberg RG, Lahoutte T. Plasma protein binding of luciferase substrates influences sensitivity and accuracy of bioluminescence imaging. Mol Imaging Biol 2011; 13: 59-66.

Keyaerts M., et al. (2008) Dynamic bioluminescence imaging for quantitative tumour burden assessment using IV or IP administration of D: -luciferin: effect on intensity, time kinetics and repeatability of photon emission. Eur J Nucl Med Mol Imaging 35: 999-1007.

  • Therapy response in oncology

Bauwens M, Wimana L, Keyaerts M, Peleman C, Lahoutte T, Kersemans K, Snykers S, Vinken M, Mertens J, Bossuyt A. Preliminary in vivo evaluation of [131I]-2-iodo-D-phenylalanine as a potential radionuclide therapeutic agent in R1M-fluc rhabdomyosarcoma tumor-bearing NuNu mice using bioluminescent imaging. Cancer Biother Radiopharm 2010; 25: 225-31.

Gainkam LO, Huang L, Caveliers V, Keyaerts M, Hernot S, Vaneycken I, Vanhove C, Revets H, De Baetselier P, Lahoutte T. Comparison of the biodistribution and tumor targeting of two 99mTc-labeled anti-EGFR nanobodies in mice, using pinhole SPECT/micro-CT. J Nucl Med 2008; 49: 788-95.

  • Stem cell and Islet cell graft monitoring

Baeyens L, Bonne S, Bos T, Rooman I, Peleman C, Lahoutte T, German M, Heimberg H, Bouwens L. Notch signaling as gatekeeper of rat acinar-to-beta-cell conversion in vitro. Gastroenterology 2009; 136: 1750-60 e13.

Coppens V, Heremans Y, Leuckx G, Suenens K, Jacobs-Tulleneers-Thevissen D, Verdonck K, Lahoutte T, Luttin A, Heimberg H, De Leu N. Human blood outgrowth endothelial cells improve islet survival and function when co-transplanted in a mouse model of diabetes. Diabetologia 2013; 56(2): 382-90.

  • DC vaccination

Van Lint S, Govaerts C, Maenhout S, Goethals L, Disy A, Benteyn D, Pen J, Bonehill A, Heirman C, Breckpot K, Thielemans K. Precilinical evaluation of TriMix and antigen mRNA-based antitumor therapy. Cancer Research 2012; 72(7): 1661-71.

Breckpot K, Escors D, Arce F, Lopes L, Karwacz K, Van Lint S, Keyaerts M, Collins M. HIV-1 lentiviral vector immunogenicity is mediated by Toll-like receptor 3 (TLR3) and TLR7. J Virol. 2010 Jun;84(11):5627-36. Epub 2010 Mar 17.

Emeagi PU, Van Lint S, Goyvaerts C, Maenhout S, Cauwels A, McNeish IA, Bos T, Heirman C, Thielemans K, Aerts JL, Breckpot K. Proinflammatory characteristics of SMAC/DIABLO-induced cell death in antitumor therapy. Cancer Research 2012; 72(6) 134-52.

Dr. Marleen Keyaerts
Labo ICMI: 0032 (0)2 477 49 91
UZ Brussel: 0032 (0)2 477 50 20