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Microfluorometric measurements of the chromatin texture in mammalian cells in different states of proliferation

Confluent L-929 mouse fibroblasts were stimulated after 11 days of nutritional deprivation by fresh medium with serum to resume proliferation in synchronized manner. At different time intervals after stimulation microscopic preparations of the cells were made with the Acriflavine-Feulgen-SITS method. The intensity distributions of the two fluorochromes were scanned across the cell nuclei in each 59 rows and columns. Data were gray level displayed and evaluated interactively. The fluorescence intensities in different cell cycle phases, monitored by flow cytometry, indicated a linear increase of nucleolar proteins from the stimulation until the next mitosis, whereas the DNA amount only increased during the S- phase. The width of the distributions revealed larger differences in fluorescence intensities shortly after the stimulation than during the S-phase with more homogeneously stained chromatin. With this procedure subpopulations of stimulated cells and of those, which did not proliferate, could be determined. The results confirm that scanning microfluorometry and evaluation of the intensity distributions yield interesting information about metabolic processes in cells and allow differentiation of subpopulations.