4.5. Giemsa Staining for the Detection of Morphological Features of Apoptosis

Giemsa stain was used to assess the morphological changes of PC-3 cells, according to the standard Giemsa procedure established by the supplier (Solarbio, Beijng, China). Briefly, PC-3 cells were plated at 3 × 10³ cells/well into a 96-well plates. PC-3 cells after treatment with 12n for 48 h were fixed with methanol (75%) for 10 min at room temperature. Then, the cells were stained with Giemsa (diluted with phosphate buffer), observed, and photographed under the microscope at 200 × (Olympus Corporation, Tokyo, Japan ) [42]. PC-3 cell lines that did not deal with the compound 12n were used as the control.