An image analysis system, assigned different pseudocolors to different types of immunolabeled cells, allowed us to make a montage from two images of the respective types of cells. This system was therefore used for simultaneous identification of two or more types of immunolabeled cells in the sheep anterior pituitary. Morphometry--including a neighboring proportion defined as the probability of a cell type adjoining other cell types--was performed. We also conducted a simulation of an artificial cell mass with an image analyzer to evaluate the effects of cell populations on the neighboring proportion. Simulation analysis showed that the predominant cell type tended to have a higher neighboring proportion, while rarer cell types had lower proportions according to their small population density. In the sheep pituitary gland, the neighboring proportions against PRL-, GH-immunolabeled cells were high (about 65% and 55%, respectively), according to their large populations. The neighboring proportion of LH beta-immunolabeled cells to the same type of cells was lower (11%) than that against other types of cells. It was thus suggested that LH cells were scattered throughout the anterior lobe. The neighboring proportion of ACTH-immunolabeled cells to the same type of cells was somewhat higher, but that of ACTH cells to PRL cells was low (52%). Accordingly, this cell type was often distributed in clusters. These quantitative results confirmed the topographical characteristics of secretory cells deduced from visual observation. In addition, a low topographical affinity between PRL and ACTH cells was indicated.