Figure 6

Comparison of caspase-3 expression and nuclear morphology using ImageJ. The caspase-3 expression of each individual cell after staurosporine-incubation was quantified and compared with the measurements of nuclear morphology to explore whether nuclear shape is correlated with apoptosis. (A) Photomicrograph showing staurosporine-incubated caspase-3+ (red) cells with DAPI-stained nuclei (blue). (B) Binary and watershed DAPI image obtained as described in Figure 3. By including the “Add to Manager” function, regions of interests (ROIs) (green) were created by ImageJ around each DAPI-stained nuclei. ImageJ enlarged the ROIs by 15 pixels to enclose most of the cell body surrounding the nucleus. (C) The ROIs (green), defining the area of fluorescence measurement, were converted to 16-bit gray scale caspase-3 images to measure the expression of caspase-3 in each cell after background fluorescence was removed by the ImageJ “Subtract Background” function. Caspase-3 expression in each cell was thereafter compared with nuclear morphology measurements. Original magnification: ×200.