The evaluation of cell nuclei staining parameters regarding the reproducibility of fluorescence measurements

The present methodological study is an attempt to optimize the staining parameters for a quantitative DNA determination of fluorescent cells. The application of pure dyes and a precise control of the staining procedure are preliminary conditions which have to be fulfilled, because the reproducibility of measurements is in this connection the most important criterion for a quantitative DNA related analysis of cell population. Chicken erythrocytes and isolated nuclei were applied as biological test objects. The staining procedure with acridine derivatives (acriflavine, proflavine, rivanol) was performed in accordance with the fluorescence-Feulgen reaction. The influence of staining parameters, such as (1) pH and (2) dye concentration of the staining solution, were evaluated regarding the spectral behaviour, the total fluorescence intensity, and the reproducibility of results.