A quantitative study of silver-stained NORs in different segments of the normal human colorectal crypt

Quantification of silver-stained nucleolar organiser regions (AgNORs) in paraffin sections may provide clues about the proliferation and differentiation in normal and neoplastic tissues. The aim of the present investigation was to determine whether AgNOR quantification could provide useful data about proliferation in the different segments of the normal human colorectal crypt. Samples of histologically 'normal' large intestine (n = 8) were obtained from colorectal cancer resections at a distance of > 5 cm from the tumour margins and were routinely processed for paraffin embedding using strictly standardised procedures. Sections were cut and stained with a one-stage silver colloid impregnation technique. The longitudinally sectioned crypts were divided into proliferative (P), intermediate (I) and surface (S) segments using strict criteria. Clearly defined AgNORs, which appeared as black dots within the nuclear profile, were quantified from each segment for volume density (Vv) and number per unit area (NA) estimates using traditional point-counting techniques. A 1-way analysis of variance followed by Scheffe's test indicated significant progressive reductions of AgNOR Vv and NA from P to S segments. Our data suggest that both volume and frequency of AgNORs may be related to cellular proliferation since both parameters are highest in the P segment. The further exploitation of stereological tools in conjunction with AgNOR staining may be valuable in assessing normal differentiation and proliferation patterns and in predicting the biological behaviour of neoplastic tissues in which increased proliferation is a feature.