Quantitation of smooth muscle proliferation in cultured aorta. A color image analysis method for the Macintosh

Color image analysis was used to assess proliferative changes in smooth muscle cells from cultured segments of rabbit aortas. Proliferating cells were labeled with bromodeoxyuridine (BrdU) and visualized by immunohistochemical staining of histologic sections. A Macintosh IIfx computer with a Data Translation digitizer board, Javelin color camera and a color-enhanced version of National Institutes of Health Image 1.31 image analysis software (ColorImage 1.31) was used to acquire red, green and blue (RGB)-filtered grayscale images from microscopic slides of control and treated aortas. The BrdU-labeled (brown) and nonlabeled, hematoxylin (blue)-stained nuclei were identified on the RGB gray-scale images using a thresholding technique and sampled for nuclear number and area. An increase in the number of BrdU-labeled nuclei in the region of experimental perturbation was demonstrated by this semiautomated method. Thus, this Macintosh-based color image analysis method proved to be effective in rapidly quantitating immunohistochemically defined smooth muscle proliferation in microscopic tissues.