A morphometric study of histological variations during cellular differentiation of normal human colorectal epithelium

Abstract

Quantifiable variations existing in the colorectal crypt during cellular differentiation were detected by using simple computer-aided morphometric techniques applied to routinely prepared H&E stained and semithin toluidine blue stained sections of normal colonic mucosa. Generally, most of the morphometric parameters including nuclear volume, nuclear volume weighted mean volume, cytoplasmic volume, cellular volume, nuclear axial ratio (a/b), mean nuclear diameter, nuclear shape factor (NSF) and nuclear maximum angle (Agmax) showed an increasing trend between basal and surface segments. Conversely, the nuclear-cytoplasmic (N/C) ratio, NSF and nuclear circularity index (NCI) decreased between these segments. Epithelial cells in the basal segment had the highest N/C ratio and the lowest cell volume due to their low volumes of cytoplasm. However, substantial increases of cytoplasmic volume occurred in the intermediate segment, thereby expanding the cell volume to 136% of that of the basal segment cell. Data for a/b, NSF, NCI and Agmax suggest that the epithelial nuclei were more ellipsoidal in shape and were aligned more perpendicular to the basement membrane as they reached the surface epithelium. Numerical densities for epithelial cell nuclei were highest in the basal segment, indicating more nuclear profiles at this region per unit area or volume. This also suggested that the basal segment was the active proliferating zone. Such observations agree with previously reported cell kinetic and autoradiographic studies.