Figure 2.
Automated single cell measurement capability of p-BNC assay. In the raw image (A), individual cell outlines were identified by applying adaptive thresholds to DAPI and Phalloidin channels for each image (B) and used to create regions of interest (ROIs) specific to each cell (C &D). These ROIs were then analyzed through automated custom macros (ImageJ, Matlab) to obtain cytomorphometric and molecular data at a single-cell level. Measurements up to 20 parameters (i.e. area, mean value, standard deviation, modal value, centroid, center of mass, perimeter, bounding rectangle, fit ellipse, circularity, Feret’s diameter, median, integrated density) are made for each color (Red, Green, Blue) separately for the cytoplasm and the nucleus.
