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Proceedings Paper

Time-lapse microscopy of lung endothelial cells under hypoxia
Author(s): Shima Mehrvar; Zahra Ghanian; Ganesh Kondouri; Amadou S. Camara; Mahsa Ranji
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Paper Abstract

Objective: This study utilizes fluorescence microscopy to assess the effect of the oxygen tension on the production of reactive oxygen species (ROS) in mitochondria of fetal pulmonary artery endothelial cells (FPAECs). Introduction: Hypoxia is a severe oxygen stress, which mostly causes irreversible injury in lung cells. However, in some studies, it is reported that hypoxia decreases the severity of injuries. In this study, ROS production level was examined in hypoxic FPAECs treated with pentachlorophenol (PCP, uncoupler). This work was accomplished by monitoring and quantifying the changes in the level of the produced ROS in hypoxic cells before and after PCP treatment. Materials and methods: The dynamic of the mitochondrial ROS production in two groups of FPAECs was measured over time using time-lapse microscopy. For the first group, cells were incubated in 3% hypoxic condition for 2 hours and then continuously were exposed to hypoxic condition for imaging as well. For the second group, cells were incubated in normal oxygen condition. Time lapse images of the cells loaded with Mito-SOX (ROS indicator) were acquired, and the red fluorescence intensity profile of the cells was calculated. Changes in the level of the fluorescence intensity profile while they are treated with PCP indicates the dynamics of the ROS level. Results: The intensity profiles of the PCP-treated cells in the first group showed 47% lower ROS production rate than the PCP-treated cells in the second group. Conclusion: Time lapse microscopy revealed that hypoxic cells have lower ROS generation while treated with PCP. Therefore, this result suggests that hypoxia decreased electron transport chain activity in uncoupled chain.

Paper Details

Date Published: 16 February 2017
PDF: 4 pages
Proc. SPIE 10068, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV, 1006822 (16 February 2017); doi: 10.1117/12.2254866
Show Author Affiliations
Shima Mehrvar, Univ. of Wisconsin-Milwaukee (United States)
Zahra Ghanian, Univ. of Wisconsin-Milwaukee (United States)
Ganesh Kondouri, Medical College of Wisconsin (United States)
Amadou S. Camara, Medical College of Wisconsin (United States)
Mahsa Ranji, Univ. of Wisconsin-Milwaukee (United States)

Published in SPIE Proceedings Vol. 10068:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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