Abstract
Purpose
This study describes an imaging strategy based on glow stick chemistry to non-invasively image oxidative stress and reactive oxygen species (ROS) production in living animals.
Procedures
Upon stimulation, phagocytes produce toxic levels of ROS to kill engulfed microorganisms. The mitochondrial respiratory chain continually generates low levels of superoxide (O2·−) that serve as a source for generation of downstream ROS, which function as regulatory signaling intermediaries. A ROS-reacting substrate, 2-methyl-6-[4-methoxyphenyl]-3,7-dihydroimidazo[1,2-a]pyrazin-3-one hydrochloride, was used as the chemical energy donor for generating energy transfer luminescence in phagosomes and mitochondria.
Results
Using targeted photoluminescent dyes with specific subcellular localization that serve as chemical energy recipients, our imaging data demonstrate proof-of-concept for using glow stick chemistry to visualize ROS production associated with phagocytosis and mitochondrial respiration in living mice.
Conclusions
Glow stick imaging is a complementary hybrid of chemiluminescence and photoluminescence imaging, capable of generating red or far-red emission for deep tissue imaging.
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Acknowledgments
This work was supported by the Nancy Lurie Marks Foundation. We thank Kin K. Wong for his assistance in preparing this report. We thank Dr. Lisa Cameron at DFCI for providing technical support of microscopic imaging. We thank Dr. Nancy E. Kohl and Dr. Annick Van den Abbeele for critical reading and insightful discussion.
Conflict of Interest
D.B., T.T., and A.L.K. declare no conflict of interest in this study. J.–C.T. submitted a patent application based on this study (DFCI).
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Tseng, JC., Bailey, D., Tupper, T. et al. Using Glow Stick Chemistry for Biological Imaging. Mol Imaging Biol 16, 478–487 (2014). https://doi.org/10.1007/s11307-014-0721-8
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DOI: https://doi.org/10.1007/s11307-014-0721-8