1. Time-lapse Imaging of Oxidative Stress in vitro 1 by: Zahra Ghanian Advisor Dr. Mahsa Ranji 04/25/2015

2. Electron Transport Chain 2  Aim: Quantify the dynamic of ROS by time lapse microscopy  Modulate ROS by metabolic perturbation  Experimental protocol baseline imagingMitoSOX imagingimaging 10 min 20 min 60 min uncoupler or inhibitorMitoSOX

3. 3 OS production dynamics  Sigmoidal fit: control cells  Exponential fit: treated cells -aTb PCP17.2435.1518.46 ROT12.8871.9513.34 AA11.0542.1911.62 KCN15.8389.1516.26 c τ d CTRL-18.719.9620.77

4. Results and Conclusion 4  Right after addition of agent, OS production significantly increased compared to control  Pulmonary hypertension is associated with greater OS production rate  Addition of KCN to PCP-treated PAEC cells resulted in 66.15% significant decrease in OS production  presented method is able to partition and quantify the dynamics of superoxide production from different ETC complexes at different stress conditions of injury and treatment.  We have developed a new methodology and protocol establish a diagnostic tool for early detection of different diseases and a correlation between predicted target organ damage and changes in the ROS levels. agent Slope ratio PCP12.05 ± 2.3 ROT3.6 ± 0.4 AA5.7 ± 0.6 KCN3.7 ± 0.6 Slope of the models at t=30 min