Darzynkiewicz Z, Traganos F, Sharpless T, Melamed M R
J Histochem Cytochem. 1977 Jul;25(7):875-80. doi: 10.1177/25.7.70457.
Cells in mitosis may be distinguished from interphase cells based on difference in chromatin structure as revealed by two different methods of staining with acridine orange. In the first method, cells are heated and then stained at neutral pH; the difference in stainability between mitotic and interphase cells reflects the difference in the extent of deoxyribonucleic acid denatured by heat in these cells. At a given temperature the deoxyribonucleic acid of the mitotic cell appears to be more extensively denatured than that of the interphase cell. In the second method, cells are treated with buffer at pH 1.5 (1.3 to 1.9) and then stained at pH 2.6 (2.3 to 2.9). The mechanisms involved in the differential stainability of interphase versus mitotic cells at that low pH are currently under investigation. In both methods, in addition to enumerating cells in mitosis, it is possible to quantitate cells in G1, S and G2 phases of the cell cycle.
通过吖啶橙的两种不同染色方法所揭示的染色质结构差异,可以区分处于有丝分裂期的细胞和间期细胞。在第一种方法中,细胞先受热然后在中性pH下染色;有丝分裂期细胞和间期细胞在染色性上的差异反映了这些细胞中因受热而变性的脱氧核糖核酸程度的差异。在给定温度下,有丝分裂期细胞的脱氧核糖核酸似乎比间期细胞的变性程度更高。在第二种方法中,细胞用pH 1.5(1.3至1.9)的缓冲液处理,然后在pH 2.6(2.3至2.9)下染色。目前正在研究在该低pH下间期细胞与有丝分裂期细胞染色性差异所涉及的机制。在这两种方法中,除了对有丝分裂期的细胞进行计数外,还可以对细胞周期G1、S和G2期的细胞进行定量。