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Detection of autophagic cells by acridine orange staining

DH Dongmei Han
XW Xingwei Wu
LL Libin Liu
WS Wanting Shu
ZH Zhenping Huang
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As a marker of autophagy, the volume of the cellular acidic compartment was visualized by acridine orange (AO) staining22. AO (Solarbio, Beijing, China), a fluorochrome, has membrane permeability that can penetrate into acidic organelles, such as autophagy- lysosomes. Autophagic cells stained by AO can be observed to have red and yellow spots, known as acidic bodies, by a fluorescence microscope. The AO staining solution (1 mg/ml) is used as the storage solution, and should be diluted to the appropriate concentration (8.5~17 μg/ml) before use. Staining was allowed to proceed for 15~20 min at room temperature in the dark. The ARPE-19 cells stained by AO were observed by a fluorescence microscope (excitation filter wavelength 488 nm, blocking filter wavelength 515 nm) and images were taken.

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