Image analysis of DNA fragmentation and loss in V79 cells under apoptosis

Abstract

Nuclear image analysis of Feulgen-stained V79 fibroblasts after three days in culture was used to discriminate apoptotic cells and cells suspected to be undergoing apoptosis from control cells based on parameters such as the Feulgen-DNA content, degree of chromatin condensation and nuclear areas, in association with visual morphology. The fibroblasts were initially plated at a density of 10(5) cells/ml and incubated under optimal culture conditions without subculturing. Following confluency, the cells underwent contact inhibition apoptosis. Image analysis revealed three nuclear phenotypes which were defined in terms of their morphological characteristics and levels of chromatin condensation. A decrease in the amount of Feulgen-DNA was detected in apoptotic cells and in cells suspected of undergoing apoptosis. This decrease was assumed to indicate DNA loss. Image analysis procedures may therefore provide a useful tool for discriminating cells in the early stages of apoptosis.