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CYTOFLUOROMETRIC QUANTITATION OF ACRIDINE ORANGE UPTAKE BY CULTURED CELLS
Author(s) -
RUNDQUIST I.,
OLSSON M.,
BRUNK U.
Publication year - 1984
Publication title -
acta pathologica microbiologica scandinavica series a :pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0108-0164
DOI - 10.1111/j.1699-0463.1984.tb04408.x
Subject(s) - acridine orange , fluorescence , staining , population , fluorescence microscope , microbiology and biotechnology , chemistry , cell , fluorescent staining , biology , biophysics , biochemistry , genetics , physics , demography , quantum mechanics , sociology
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 (> 10 g/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.