13 February 2007 Optical imaging the redox status change during cell apoptosis
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Abstract
Many cellular events involve the alteration in redox equilibrium, globally or locally. In many cases, excessive reactive oxygen species (ROS) production is the underlying cause. Several green fluoresecence protein based indicators are constructed to measure redox status in cells, e.g, rxYFP and roGFPs, which allow real time detection. reduction and oxidization-sensitive GFP (RoGFPs) are more useful due to ratiometric variation by excitation, making the measurement more accurate. Utilizing one of those roGFPs called roGFP1, we establish a mitochondrial redox state probing platform in HeLa cells with laser scan confocal microscopy (LSCM) as detection system. Control experiments confirmed that our platform could produce stable ratiometric values, which made the data more accurately reflect the real environmental changes of redox status that roGFP1 probed. Using exogenous H2O2 and DTT, we evaluated the reactivity and reversibility of roGFP1. The minimal hydrogen peroxide concentration that roGFP1 could show detectable ratiometric changes in our system was about 200&mgr;M. Preliminarily applying our platform to exploring the redox status during apoptosis, we observed an increase in ratiometric, suggesting an excessive ROS production.
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Ting Su, Ting Su, Zhihong Zhang, Zhihong Zhang, Juqiang Lin, Juqiang Lin, Qingming Luo, Qingming Luo, } "Optical imaging the redox status change during cell apoptosis", Proc. SPIE 6438, Biophotonics and Immune Responses II, 64380K (13 February 2007); doi: 10.1117/12.701654; https://doi.org/10.1117/12.701654
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