A fluorescence-based approach to probing the immediate/early molecular mechanisms of photobiomodulation in vitro

Nathaniel J. Pope; Michael L. Denton

doi:10.1117/12.2508961

7 March 2019 A fluorescence-based approach to probing the immediate/early molecular mechanisms of photobiomodulation in vitro

Nathaniel J. Pope, Michael L. Denton

Author Affiliations +

Proceedings Volume 10861, Mechanisms of Photobiomodulation Therapy XIV; 108610F (2019) https://doi.org/10.1117/12.2508961
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

Photobiomodulation (PBM) describes the use of low levels of visible to near infrared light to induce positive biological effects. While there is a growing body of research demonstrating therapeutic and protective benefits of PBM, the underlying molecular mechanisms remain poorly understood. The putative chromophore for PBM effects is cytochrome c oxidase, also known as complex IV (C-IV) of the mitochondrial electron transport chain. It is believed that via action on C-IV, light absorption initiates a cascade of events involving nitric oxide (NO), reactive oxygen species (ROS), and increased mitochondrial function, leading to improved cellular robustness. However, little is known about the mechanisms by which these signaling pathways are initiated and how they establish the observed beneficial outcomes. We have conducted in vitro analyses of living human cells to probe changes in NO, ROS, and mitochondrial function, particularly focusing on the immediate and early effects of light exposure. This analysis utilizes a novel multi-channel laser assembly, integrated with a fluorescence microscope, and collimated for a sub-field-of-view beam diameter. By combining this apparatus with specific vital stains, we can visualize production of NO, ROS, and changes in mitochondrial morphology and dynamics in real-time, during and immediately following light exposure. Additionally, we are able to compare these effects to unexposed cells within the same field-of-view. Moving forward, future studies will examine additional vital stains in cells and isolated mitochondria, allowing us to distinguish direct mitochondrial effects from those requiring cellular feedback. In summary, this research provides a new window into the molecular underpinnings of the PBM response.

Citation Download Citation

Nathaniel J. Pope and Michael L. Denton "A fluorescence-based approach to probing the immediate/early molecular mechanisms of photobiomodulation in vitro", Proc. SPIE 10861, Mechanisms of Photobiomodulation Therapy XIV, 108610F (7 March 2019); https://doi.org/10.1117/12.2508961

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