Gregoire M, Hernandez-Verdun D, Bouteille M
Exp Cell Res. 1984 May;152(1):38-46. doi: 10.1016/0014-4827(84)90228-3.
Chromatin distribution was visualized in living cells with the selective DNA fluorochrome Hoechst 33342. This dye was shown to be non-toxic on the rat kangaroo PTO cell line by measuring the labelled cell growth rate. The aim of this work was firstly to visualize chromatin distribution without fixation or dehydration and secondly to demonstrate that quantitative determination of DNA content was possible under these non-toxic labelling conditions. During interphase, condensed, decondensed and thin network chromatin configurations were visualized. In nucleolar regions the fluorochrome revealed well-defined chromocentres. During mitosis, fluorescent chromosome banding was observed in vital conditions and chromocentres on fixed chromosomes. Chromatin segregation was visualized after micronucleation, which induced chromosomal set distribution in individual micronuclei. By this means, we demonstrated that the chromocentres observed in interphase nuclei were part of nuclear organizer region (NOR)-bearing chromosomes. This vital staining of chromatin was shown to be compatible with the quantitative determination of DNA content, both in living PTO cells and in isolated nuclei.
利用选择性DNA荧光染料Hoechst 33342在活细胞中观察染色质分布。通过测量标记细胞的生长速率，表明该染料对大鼠袋鼠PTO细胞系无毒。这项工作的目的，一是在不进行固定或脱水的情况下观察染色质分布，二是证明在这些无毒标记条件下可以对DNA含量进行定量测定。在间期，可以观察到凝聚、解聚和细网状的染色质构型。在核仁区域，荧光染料显示出清晰的染色中心。在有丝分裂期间，在活体条件下观察到荧光染色体带，以及固定染色体上的染色中心。微核形成后观察到染色质分离，微核形成导致单个微核中的染色体组分布。通过这种方法，我们证明了在间期核中观察到的染色中心是携带核仁组织区（NOR）的染色体的一部分。这种对染色质的活体染色被证明与在活的PTO细胞和分离的细胞核中对DNA含量的定量测定是兼容的。
