Quantitation with an automated image analyzer of nuclearcytoplasmic changes induced by hydrocortisone in bladder...

Thirty-eight urinary bladders from Fischer rats were organ cultured for 7 days in Ham's Medium F12 with varying concentrations of hydrocortisone added. Measurements of cell numbers and relative areas of nuclei and cytoplasm (N/C ratios) were made with an automated image analyzer on 1.5-micronm-thick plastic sections of the tissues. Epithelia cultured in the absence of hydrocortisone became hyperplastic and dysplastic and contained approximately 2.5 times as many cells per unit length as uncultured controls. N/C ratios were increased by over 50%, caused mainly by a decrease in the cytoplasmic area. Concentrations of hydrocortisone below 10(-7) M had no significant effect on the hyperplasia or N/C ratios. Increasing concentrations above 10(-7) M resulted in progressive decreases of both the number of cells per unit length and the N/C ratios. Epithelia cultured in 2 X 10(-5) M hydrocortisone were not significantly different from uncultured controls. N/C ratios from 85 fields with a wide range of values were quanititated both with the image analyzer and a manual point-counting technique. Linear regression analysis revealed a close linear relationship and a correlation coefficient of 0.922. The analyzer provided an efficient, accurate tool for quantitating morphological observations.