Acridine Orange - CAS 65-61-2 - Calbiochem

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For viability determination at each time point, AO/PI double staining was performed. AO is a stain that is permeable to viable cells and can stain the cell’s DNA directly. This dye emits a green fluorescence once it is excited. PI, on the other hand, is a dye that is impermeable to viable cells. It can bind to DNA only when the cells are dead; it emits a red-orange fluorescence instead

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For viability determination at each time point, AO/PI double staining was performed. AO is a stain that is permeable to viable cells and can stain the cell’s DNA directly. This dye emits a green fluorescence once it is excited. PI, on the other hand, is a dye that is impermeable to viable cells. It can bind to DNA only when the cells are dead; it emits a red-orange fluorescence instead

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For viability determination at each time point, AO/PI double staining was performed. AO is a stain that is permeable to viable cells and can stain the cell’s DNA directly. This dye emits a green fluorescence once it is excited. PI, on the other hand, is a dye that is impermeable to viable cells. It can bind to DNA only when the cells are dead; it emits a red-orange fluorescence instead. After removing the solutions and culture medium, cells were washed twice with PBS. Cells were stained with an equal volume combination of AO and PI for 20 seconds and washed twice with PBS for removing dyes. The stained sample was viewed under a fluorescent microscope. After AO/PI staining, cells were counted according to the method described in previous studies to determine the potential of different cardioplegic solutions to induce apoptosis and necrosis on other cells. According to the previous studies, the criteria are as follows: uniform green nucleus with organized structure, intact plasma membrane, orange or green cytoplasm, viable cells 1), bright green areas of chromatin condensation in the nucleus, early apoptosis 2), dense orange areas of chromatin condensation, late apoptosis 3), orange intact nucleus, and necrosis 4). For each test group, three fluorescence microscope images with a magnification of 320 were analyzed by examining and counting cells manually. The quantification of apoptotic and necrotic cells was calculated according to the following equations.

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