*4.8. Evaluation of Nuclear Morphology*

Nuclear fragmentation and chromatin condensation of the cancer cells were evaluated by using the nuclear staining dye Hoechst 33342 (10 μg mL−1) and via the double staining method using acridine

orange/ethidium bromide (100 μg mL−1). Experiments were carried out, according to Fernando et al. (2018) [21]. Briefly, 24 h pre-seeded cells were treated with di fferent concentrations of the samples and incubated for 24 h. The fluorescence dyes were then applied to the wells and incubated for 10 min. The images were visualized using a fluorescence microscope with a CoolSNAP-Pro color digital camera.
