Evos fl 2 auto 2 cell imaging system

The morphological changes of MCF-7 cell death induced by Ph₃SnL1 and cisplatin were studied by AO (acridine orange) [41 (link)] and DAPI (4′,6-diamidino-2-phenylindole) [42 (link)] staining, and examined under a fluorescence microscope. Acridine orange can cross the cell membrane and viable and early apoptotic cells can be detected. Chromatin condensation, seen as dense green areas, or membrane blebbing, both appearing in apoptosis, is easily proven by AO staining [40 (link)]. After 24 h of treatment (IC₅₀ and 2 × IC₅₀ concentrations) the cells were stained with acridine orange (3 μg/mL AO in PBS), and observed with a fluorescence microscope (Fluorescence microscope-Invitrogen EVOS FL 2 Auto 2 Cell Imaging System). On the other hand, for the DAPI assay 0.5 mL of Triton (0.1% in PBS) was added to the treated cells after the fixing the cells with 0.5 mL of 4% paraformaldehyde (PFA) for 8 min. Thereafter, 0.5 mL of DAPI solution was used to stain the cells for 5 min in the dark [42 (link)]. The morphological characteristics were visualized under a fluorescence microscope (Fluorescence microscope-Invitrogen EVOS FL 2 Auto 2 Cell Imaging System).