Investigation of reaction mechanisms of cytochrome c and mitochondria with transient absorption spectroscopy

Sean P. O'Connor; Joshua W. Lalonde; John M. Rickman; Gary D. Noojin; Nathaniel J. Pope; Michael L. Denton; Marlan O. Scully; Vladislav V. Yakovlev

doi:10.1117/12.2508170

7 March 2019 Investigation of reaction mechanisms of cytochrome c and mitochondria with transient absorption spectroscopy

Sean P. O'Connor, Joshua W. Lalonde, John M. Rickman, Gary D. Noojin, Nathaniel J. Pope, Michael L. Denton, Marlan O. Scully, Vladislav V. Yakovlev

Author Affiliations +

Proceedings Volume 10861, Mechanisms of Photobiomodulation Therapy XIV; 108610E (2019) https://doi.org/10.1117/12.2508170
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

We performed Transient Absorption Spectroscopy (TAS) on samples of cytochrome c in its reduced and oxidized states as well as mitochondria, and we use kinetic analysis to determine the decay rates of the transients. We also tested the samples following red light exposures to determine if there were any changes in transient decay rates. Mitochondria were isolated from hTERT-RPE1 cells and were tested in a glutamate buffer solution while cytochrome c was tested in a Sodium Phosphate solution (Na₂PO₄). Femtosecond TAS was performed with a pump pulse with wavelength centered at 418 nm and a supercontinuum probe pulse with a spectrum between 440 and 760 nm. Red light illuminations were performed with a 635 nm continuous wave light source at an intensity of 1.6 mW/cm² for 0.5 hours. We find transients and kinetic decay rates for reduced and oxidized cytochrome c that are consistent with previous literature. However, we do not find any transients for mitochondria under our current TAS testing parameters, and we expect that this may be due to low Signalto- Noise ratio or choice of pump wavelength.

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Sean P. O'Connor, Joshua W. Lalonde, John M. Rickman, Gary D. Noojin, Nathaniel J. Pope, Michael L. Denton, Marlan O. Scully, and Vladislav V. Yakovlev "Investigation of reaction mechanisms of cytochrome c and mitochondria with transient absorption spectroscopy", Proc. SPIE 10861, Mechanisms of Photobiomodulation Therapy XIV, 108610E (7 March 2019); https://doi.org/10.1117/12.2508170

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