Figure 5.

Heatmaps tables illustrating toxicity indicated parameters in A549 cells exposed to SiO₂NP. A549 cells growing on 96-well plates were exposed to various concentrations (0.01, 0.1, or 0.5 mg/ml) of 20 nm (A), 30 nm (B), 40 nm (C), 80 nm (D), or 400 nm (E) SiO₂NP for 1 h, 3 h, 6 h, or 24 h. Multi-parameter high content screening analysis for cell count, Lysosomal mass/pH and cell membrane permeability (MP) was performed using an automated IN Cell Analyzer 1000 microscope and IN Cell Investigator image analysis software. Data represents three independent experiments performed in triplicate samples. Heatmaps colorimetric gradient table for the A549 results span from: Dark green = lower than 15% of maximum value measured; Bright green = 30%; Yellow = 50%; Bright Orange = 60%; Dark Orange = 75%; Red = higher than 75% of maximum value. Cell viability colour gradients read as percentage of cell loss compared to normalised control (green = low cell viability loss, red = high loss) compared to normalised control. Heatmaps values are normalised using the percent of the positive controls and, Z score was calculated as described in the statistical analysis section.