chapter  15
24 Pages

- Bioluminescence Imaging

ByJuliaan R.M. van Rappard, Preston Lavinghousez, and Joseph C. Wu

BLI requires the transfection of cells or tissues with a gene cassette consisting of a selected promoter that controls the expression of the bioluminescence reporter gene. Once transfected, the expression cassette produces the luciferase enzyme, and aer the specic

Optical Techniques in Regenerative Medicine. Edited by Stephen P. Morgan, Felicity R.A.J. Rose, and Stephen J. Matcher © 2014 CRC Press/Taylor & Francis Group, LLC. ISBN: 978-1-4398-5495-2

15.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 403

15.2 Clinical and Biological Measurement Challenges the Method is Suited to Address . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 405

15.3 Technical Implementation of the Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 406 15.3.1 Bioluminescence Reporter Genes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 406 15.3.2 Instrumentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 408

15.4 Cardiovascular Disease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 408

15.5 Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 411

15.6 Transplant Immunology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 412

15.7 Central Nervous System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 413

15.8 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 414

Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 415

References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 415

reporter probe is introduced (luciferin or coelenterazine), an oxidation reaction occurs in the presence of magnesium, oxygen, and adenosine triphosphate (ATP; for luciferaseluciferin), emitting visible light photons that can be detected by a charge coupled device (CCD) camera. is reaction allows in vivo tracking of the injected cells within the subject via detecting the emitted photon light signal (Figure 15.1). In planar BLI, there is a 10-fold loss of photon density for each centimeter of tissue depth (Contag et al. 1995). us far, repeated exposure of animals to reporter probes appears to be safe and nontoxic. Due to tissue attenuation properties, the cells emitting light closer to the surface appear brighter compared to deeper sources in creating surface-weighted images (Weissleder 2001; Wu et al. 2001).