Identification of Factors Complicating Bioluminescence Imaging

Biochemistry. 2019 Mar 26;58(12):1689-1697. doi: 10.1021/acs.biochem.8b01303. Epub 2019 Mar 11.


In vivo bioluminescence imaging (BLI) has become a standard, non-invasive imaging modality for following gene expression or the fate of proteins and cells in living animals. Currently, bioluminescent reporters used in laboratories are mostly derivatives of two major luciferase families: ATP-dependent insect luciferases and ATP-independent marine luciferases. Inconsistent results of experiments using different bioluminescent reporters, such as Akaluc and Antareas2, have been reported. Herein, we re-examined the inconsistency in several experimental settings and identified the factors, such as ATP dependency, stability in serum, and molecular sizes of luciferases, that contributed to the observed differences. We expect this study will make the research community aware of these factors and facilitate more accurate interpretation of BLI data by considering the nature of each bioluminescent reporter.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Enzyme Stability
  • Genes, Reporter
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Imidazoles / metabolism
  • Luciferases / blood
  • Luciferases / genetics
  • Luciferases / metabolism*
  • Luminescent Agents / metabolism
  • Luminescent Measurements / methods*
  • Mice, Inbred BALB C
  • Neoplasms / diagnostic imaging
  • Pyrazines / metabolism
  • Tissue Distribution


  • Imidazoles
  • Luminescent Agents
  • Pyrazines
  • Adenosine Triphosphate
  • Luciferases