In vivo imaging of reactive oxygen and nitrogen species in inflammation using the luminescent probe L-012

Free Radic Biol Med. 2009 Sep 15;47(6):760-6. doi: 10.1016/j.freeradbiomed.2009.06.013. Epub 2009 Jun 17.


Production of reactive oxygen and nitrogen species (ROS/RNS) is an important part of the inflammatory response, but prolonged elevated levels of ROS/RNS as under chronic inflammation can contribute to the development of disease. Monitoring ROS/RNS in living animals is challenging due to the rapid turnover of ROS/RNS and the limited sensitivity and specificity of ROS/RNS probes. We have explored the use of the chemiluminescent probe L-012 for noninvasive imaging of ROS/RNS production during inflammation in living mice. Various inflammatory conditions were induced, and L-012-dependent luminescence was recorded with an ultrasensitive CCD camera. Strong luminescent signals were observed from different regions of the body corresponding to inflammation. The signal was reduced by administration of the SOD mimetic tempol, the NADPH oxidase inhibitor apocynin, and the inhibitor of nitric oxide synthesis L-NAME, signifying the requirement for the presence of ROS/RNS. Additionally, the L-012 signal was abolished in mice with a mutation in the Ncf1 gene, encoding a protein in the NADPH oxidase complex 2, which generates ROS/RNS during inflammation. In conclusion, L-012 is well distributed in the mouse body and mediates a strong ROS/RNS-dependent luminescent signal in vivo and is useful for monitoring the development and regulation of inflammation in living organisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetophenones / pharmacology
  • Animals
  • Cyclic N-Oxides / pharmacology
  • Diagnostic Imaging / instrumentation
  • Diagnostic Imaging / methods*
  • Inflammation
  • Luminescent Agents*
  • Luminol / analogs & derivatives*
  • Mice
  • NADPH Oxidases / antagonists & inhibitors
  • NADPH Oxidases / genetics
  • NG-Nitroarginine Methyl Ester / pharmacology
  • Nitric Oxide Synthase / antagonists & inhibitors
  • Oxidative Stress / genetics
  • Reactive Nitrogen Species / analysis*
  • Reactive Nitrogen Species / metabolism
  • Reactive Oxygen Species / analysis*
  • Reactive Oxygen Species / metabolism
  • Sensitivity and Specificity
  • Spin Labels
  • Superoxide Dismutase / antagonists & inhibitors


  • Acetophenones
  • Cyclic N-Oxides
  • Luminescent Agents
  • Reactive Nitrogen Species
  • Reactive Oxygen Species
  • Spin Labels
  • L 012
  • Luminol
  • acetovanillone
  • Nitric Oxide Synthase
  • Superoxide Dismutase
  • NADPH Oxidases
  • neutrophil cytosolic factor 1
  • tempol
  • NG-Nitroarginine Methyl Ester