ROBBINS E, MARCUS P I
J Cell Biol. 1963 Aug;18(2):237-50. doi: 10.1083/jcb.18.2.237.
The in vitro localization of acridine orange (AO) in living cells was monitored by means of fluorescence microscopy, quantitative cell viability studies, and photofluorimetric measurements following dye-cell interaction. The parameters, pH, time, dye concentration, and the metabolic state of the cell were found to exert a profound influence on the time course and distribution of staining. The parameters studied are mutually interdependent, and intracellular dye localization may be predictably altered by their appropriate manipulation. Conditions are defined whereby two morphologically distinct but physiologically interrelated reactions, namely, acridine orange particle (AOP) formation and cytoplasmic reddening (CR) may be caused, prevented, reversed, or modified. These results are explained in terms of the facilitation or inhibition of an intracytoplasmic dye-segregating mechanism, in turn affected by the rate of dye ingress and the physiological state of the cell. Whereas the accumulation of AO in AOP is compatible with cell viability, the appearance of CR is correlated with cell death. It is pointed out that meaningful interpretation of vital staining requires precise regulation of many parameters in the extracellular milieu. A scheme of cell compartmentalization with respect to AO is proposed to satisfactorily account for the effects of environmental variations on the distribution and ultimate fate of intracellular dye. The AOP are viewed as normally present acid phosphatase-positive multivesicular bodies.
通过荧光显微镜、定量细胞活力研究以及染料与细胞相互作用后的光荧光测量，监测吖啶橙（AO）在活细胞中的体外定位。发现pH值、时间、染料浓度和细胞的代谢状态等参数对染色的时间进程和分布有深远影响。所研究的参数相互依存，通过适当操作可预测地改变细胞内染料的定位。定义了一些条件，据此可引发、防止、逆转或改变两种形态上不同但生理上相关的反应，即吖啶橙颗粒（AOP）形成和细胞质变红（CR）。这些结果根据细胞质内染料分离机制的促进或抑制来解释，而这又依次受染料进入速率和细胞生理状态的影响。虽然AO在AOP中的积累与细胞活力相符，但CR的出现与细胞死亡相关。指出对活细胞染色进行有意义的解释需要精确调节细胞外环境中的许多参数。提出了一个关于AO的细胞区室化方案，以令人满意地解释环境变化对细胞内染料分布和最终命运的影响。AOP被视为正常存在的酸性磷酸酶阳性多囊泡体。
