Rundquist I, Olsson M, Brunk U
Acta Pathol Microbiol Immunol Scand A. 1984 Sep;92(5):303-9. doi: 10.1111/j.1699-0463.1984.tb04408.x.
The vacuolar accumulation of the lysosomotropic weak base acridine orange (AO) within living cells in culture was studied by cytofluorometry. Mouse peritoneal macrophages, malignant human glioma cells, and normal human glial cells were utilized. Exposure to AO resulted in granular bright red fluorescence, as well as a diffuse weak green background fluorescence. To obtain reproducible "staining" conditions, the red granular fluorescence was measured as a function of dye concentration and staining time. Exposure to high concentrations of AO (greater than 10 micrograms/ml) was found to cause cell damage in combination with markedly changed fluorescence distribution for the cell population with reduced mean fluorescence and increased variability. Granular uptake of AO was pH-dependent and almost zero at pH 5.5. AO fluorescence, as measured by cytofluorometry, was found to be roughly linear to the amount of AO present in the cells, as measured by spectrofluorometry after cell solubilization, indicating negligible fluorescence quenching. AO labelling of living cells might serve as a useful indicator of the condition of the cellular vacuolar (lysosomal) apparatus.
通过细胞荧光测定法研究了培养的活细胞内溶酶体促渗弱碱吖啶橙（AO）的液泡积累情况。使用了小鼠腹膜巨噬细胞、恶性人类胶质瘤细胞和正常人类神经胶质细胞。暴露于AO会导致颗粒状亮红色荧光以及弥漫性弱绿色背景荧光。为了获得可重复的“染色”条件，将红色颗粒荧光作为染料浓度和染色时间的函数进行测量。发现暴露于高浓度的AO（大于10微克/毫升）会导致细胞损伤，同时细胞群体的荧光分布明显改变，平均荧光降低且变异性增加。AO的颗粒摄取是pH依赖性的，在pH 5.5时几乎为零。通过细胞荧光测定法测量的AO荧光，与细胞溶解后通过荧光分光光度法测量的细胞内AO含量大致呈线性关系，表明荧光猝灭可忽略不计。活细胞的AO标记可能是细胞液泡（溶酶体）装置状况的有用指标。
