Appendix Fig. A3.

Separation of mitotic cells from interphase cell by MPM-2 labeling intensity. (a) All cells with 2N or 4N DNA content are plotted on a two-dimensional density plot, with median cytoplasmic MPM-2 pixel intensity on the y-axis and median nuclear MPM-2 pixel intensity on the x-axis. The frequency of cells falling in particular bin is indicated by its color, with black meaning no cells, then with increasing frequency from blue to green to yellow to red to white. (b) A portion of the same cells were manually classified into cell cycle stages, and the total cellular DAPI sum of pixel intensities for cells of each stage were plotted as on a scatterplot with the same axes as in (a), with spots color coded by stage (interphase, black; prophase, aqua; prometaphase, blue; metaphase, red; anaphase, yellow; cytokinesis, purple; telophase, olive). In both (a) and (b), red lines represent nuclear and cytoplasmic median MPM-2 pixel intensity thresholds that separate cells of the mitotic class above either threshold from cells of the interphase class below both thresholds. (c) The percentage of each manually determined cell cycle phase or mitotic stage (row headings) to fall into each automatic class (column headings) is shown.