Confocal microscopy analysis of ROS production in EMFs-exposed PBLM cells.

<p>PBLM cells exposed to EMFs for the indicated time were incubated with 2,7-dichlorofluorescin diacetate and analyzed by confocal microscopy as detailed in Materials and Methods. (<b>A</b>) Representative images are shown with enlargement of selected cells. (<b>B</b>) DCF-stained PBLMs were clustered as a function of the calculated area and the fluorescence intensity/cell averaged (from 50 to 100 single cell-analysis under each condition). The values are means ±SEM of 3 biological replicates. The black bars in the graph denote the size range of the two major cell subsets indicated as small and large size PBLM (S-PBLM and L-PBLM respectively). Image analysis was performed in threshold adjusted optical fields using the “particle analysis” tool of ImageJ. Conditions for statistical significant difference (i.e. P < 0.05): “a”, vs CTRL of the same size cluster; “b”, 12 h vs 5 h of the same size cluster; “c”, 20 h vs 12 h of the same size cluster; “d”, L-PBLM vs S-PBLM subjected to the same EMF-exposure times.