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Figure 3 | Particle and Fibre Toxicology

Figure 3

From: Mechanisms of oxidative stress and alterations in gene expression by Libby six-mix in human mesothelial cells

Figure 3

Determination of ROS generation via detection of DCFDA by flow cytometry and fluorescent microscopy. ROS generation in LP9/TERT-1 cells following (A) 8 h or (B) 24 h exposures to glass beads (non-pathogenic control), Libby six-mix, or 10 mM H2O2 (positive control) as determined by flow cytometric detection of DCFDA staining. White and black arrowheads indicate log shifts following exposure to H2O2 (10 mM; 20 min) and Libby six-mix (75×106 μm2/cm2), respectively. (C) DCFDA levels were also visualized using fluorescence microscopy. White arrowheads indicate increased DCFDA levels in Libby six-mix (75×106 μm2/cm2) treated cells. In the merged image, black arrowheads indicate identical areas defined by the white arrowheads, and demonstrate that the highest levels of ROS are produced in areas were Libby six-mix contacts LP9/TERT-1 cells. Surface area (x106 μm2/cm2) is represented in parentheses following mineral name. Bottom panel shows co-fluorescence of Hoechst dye (red nuclear stain) and DCFDA. Scale bar = 50 μm.

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