Bjelkenkrantz K
Histochemistry. 1983;79(2):177-91. doi: 10.1007/BF00489780.
The performance of Feulgen-acriflavine-SO2 and Hoechst 33258 for cytofluorometric ploidy determination is evaluated and compared. The fast initial fading of acriflavine-SO2 stained nuclei, with substantial reduction in fluorescence intensity within about 10 ms, is shown to be related to changes in DNA-ratios between cell types when measurements are performed using different excitation light intensities. Although low intensity excitation-, without such fading-, was used, the acriflavine-SO2 staining procedure showed large specimen to specimen variability regarding both mean fluorescence intensities and coefficients of variation (CV). The cells in such specimens could be shown to contain the same amount of chromophore using scanning absorbance measurements and the differing fluorescence values therefore probably represent variability in fluorescence yield. Specimens stained with Hoechst 33258 after RNase treatment showed very small deviations in mean fluorescence value between slides, and also much lower CV's than in acriflavine-SO2 stained specimens. The results indicate that with proper internal standards this procedure would allow the detection of deviations in DNA content in the order of 2%.
对用于细胞荧光测定倍性的福尔根-吖啶黄-SO₂和Hoechst 33258的性能进行了评估和比较。吖啶黄-SO₂染色的细胞核快速初始褪色，在约10毫秒内荧光强度大幅降低，结果表明，当使用不同激发光强度进行测量时，这与细胞类型之间的DNA比率变化有关。尽管使用了低强度激发（无此类褪色现象），但吖啶黄-SO₂染色程序在平均荧光强度和变异系数（CV）方面均显示出较大的样本间差异。使用扫描吸光度测量可证明此类样本中的细胞含有相同量的发色团，因此不同的荧光值可能代表荧光产率的差异。经核糖核酸酶处理后用Hoechst 33258染色的样本，玻片间平均荧光值的偏差非常小，且CV也比吖啶黄-SO₂染色的样本低得多。结果表明，使用适当的内标，该程序能够检测到约2%的DNA含量偏差。
